FORM 2
THE PATENTS ACT, 1970
(39 of 1970)
AND
THE PATENT RULES, 2003
COMPLETE SPECIFICATION
(See Section 10; rule 13)
Method and Apparatus for Gravity assisted High throughput Disruption of Cellular Material."
Applicant: Bench Bio Private Limited
Nationality: Indian
Address: CIO Jai Research Foundation
Near Daman Ganga Bridge, N.H.No. 8, Valvada,
District Valsad, Gujarat (India)
The following specification particularly describes the invention and the manner in which it is to be performed:-

FIELD OF INVENTION
The present invention relates to a gravity assisted high throughput method and apparatus for disruption of organic cells. More particularly, the present invention relates to a gravity assisted high throughput method and apparatus for disruption of organic cells and extraction of cellular material therefrom. .
BACKGROUND OF THE INVENTION
Advancement in the biotechnology requires the development of new and novel devices in the field of cell extraction and tissue lysis for extraction of cellular material. Although automated or semi-automated procedures have been developed for cell extraction techniques, disruption of the desired cell and extraction of the cellular material is an issue with such techniques due to grinding at very high RPMs.
It has often been seen in apparatuses specified for operation at high oscillation frequency that apart from disruption of the cell, the desired cellular material is also sheared thereby causing degradation and sometimes reducing the overall yield of the cellular materials. On the other hand, at a low oscillation frequency, the organic cell is insufficiently disrupted thereby resulting in reduced yield of the cellular materials. It has further been found that the overall yield of the cellular material by an apparatus depends on several other factors apart from oscillation frequency such as the stroke length and the time period of operation. The precise oscillation frequency, the desired stroke length and the time period of operation which all combine to offer a highest yield of the desired cellular material cannot be reasonably predicted by a person skilled in the art.
It has further been seen that 'the extent of disruption of organic cells given in a sample is dependent upon several factors such as stroke length i.e. the full length of movement of the sample well vertically, the oscillation frequency and the time period allowed for disruption. It has been found that with increasing stroke length or the oscillation frequency, the impact of the steel balls on the organic cell increases which leads to the disruption of the cell wall. Upon further increasing the stroke length or the oscillation frequency, the impact strength further increases, which may also shear the cellular

material apart from disruption of the cell whereas lower stroke length leads to an insufficient disruption of the cell. Thus, there is a need in the art for an apparatus having such a suitable stroke length and/or oscillation frequency that leads to a maximum disruption of the organic cells without shearing the cellular material thereby maximizing the yield of the cellular materials.
In some of the conventional methods used for cellular material extraction, the grinding jar of extractor performs radial oscillations in a horizontal position. However, such machines require more time of around two minutes and high oscillation frequency to disrupt the cells. These machines are also time-consuming and high oscillation frequency often ends up in shearing or disrupting the desired cellular material such as DNA as well.
It is often required to disrupt the organic cells in a frozen condition at temperatures of the ordeT of -196°C in order to preserve the cellular material to be extracted. The conventionally known machines require at least around two minutes of operating time in order to completely disrupt the organic cells for an appreciable quantity of the cellular material to be extracted. However, it has been seen that within about 20 to 30 seconds of operating time, the machine begins to warm up substantially and reaches a temperature of about -80°C, at which temperature the cellular material may begin to degrade. Thus, in order to preserve the cellular material, machines which maintain a cryogenic temperature throughout the operating time are known. However, such machines are expensive and maintenance of cryogenic temperature during the operating time is tedious and cost intensive. Thus, there is an unmet need in the art for an apparatus for the disruption of organic cells which is capable of disrupting substantially all of the organic cells present in the sample within about 20 seconds to about 30 seconds.
Another problem existing with the known conventional cell material extractors are that these machines are capable of simultaneously disrupting only a limited number of organic cell samples. There exists a need in the art for an apparatus that is capable of high throughput disruption of the cells of a large number of samples simultaneously.

Thus, there is a need in the art for a high throughput apparatus capable of simultaneously disrupting the organic cells of a large number of samples within 20 seconds to 30 seconds to afford a maximum yield of the intra-cellular materials at a desired oscillation frequency and at a desired stroke length to cause minimum shear of the desired intracellular materials.
These and other problems existing within the art are solved by way of the invention described hereinafter.
OBJECTS OF THE INVENTION
The various embodiments of the present invention may, but do not necessarily, achieve one or more of the following advantages and/or objects:
An object of the present invention is to provide a method and apparatus for simultaneous disruption of the organic cells present in a plurality of organic samples which overcomes the above disadvantages existing in the art.
Another object of the present invention is to provide a method and apparatus for simultaneous disruption of organic cells at a sufficient oscillation frequency that is sufficient to substantially disrupt the organic samples without shearing the desired intracellular material.
Yet another object of the present invention is to provide an apparatus having a desired stroke length that allows sufficient disruption of organic cells in a sample without shearing the desired intracellular material.
Another object of the present invention is to provide an apparatus for cellular disruption of a sample which reduces the grinding time of the sample to ensure complete disruption of the sample within about 20 seconds to about 30 seconds.
Yet another object of the present invention is to provide a method for apparatus for cellular disruption of a sample that allows maximum yield of the intracellular material.

Another object of the present invention is to provide an apparatus for simultaneous cellular disruption in a large number of samples wherein the samples perform a vertical oscillation to enable gravity assisted disruption of the organic cells.
Yet another object of the present invention is to provide a method and apparatus for simultaneous cellular disruption in a large number of samples which eliminates the need for the disruption to be carried out under cryogenic conditions.
Another object of the present invention is to provide a high throughput apparatus capable of simultaneously disrupting the organic cells of a large number of samples within 20 seconds to 30 seconds to afford a maximum yield of the intra-cellular materials at a desired oscillation frequency and at a desired stroke length to cause minimum shear of the desired intracellular material.
Yet another object of the present invention is to provide a cost-effective and convenient method of extraction of desired intracellular materials by rapid disruption of an organic cells.
These and other advantages may be realized by reference to the remaining portions of the specification and abstract.
SUMMARY OF THE INVENTION
An apparatus for gravity assisted high throughput disruption of a plurality of organic cellular material comprising:
(a) a motor;
(b) a rotatable motor shaft connected to said motor and having one end protruding out of said motor;
(c) a second rotatable shaft substantially housed within an apparatus casing and being rotatably connected to the motor shaft;
(d) a crank disposed on said second rotatable shaft which receives rotatory motion from the second shaft and converts the received rotatory motion into reciprocating motion of a connecting rod connected thereto through a crank pin;

(e) a crank pin disposed on said crank and adapted to accommodate a connecting rod;
(f) a connecting rod being connected to said crank through said crank pin at a first end such that the reciprocating motion generated by the crank is received by the connecting rod through the crank pin and transmitted to a piston shaft connected at second end of said connecting rod through a provided second crank pin;
(g) a piston shaft having a first end connected to said connecting rod through the second crank pin and a second end protruding out of said apparatus casing;
(h) a platform placed on said protruding end of the piston shaft and adapted to hold a plurality of sample boxes;
(i) a clamping plate adapted to cover a plurality of sample boxes placed on the platform, said clamping plate having a plurality of apertures, each said aperture being threaded internally; and
(j) a plurality of tie rods being externally threaded through a substantial length thereof such that said internal thread provided on said plurality of clamping plate apertures engages with said external thread provided on said tie rods to hold said sample boxes firmly in place such that a reciprocating motion of said piston shaft causes a vertical motion of the sample box, each said sample box having a plurality of wells provided therein, each said well accommodating a vial containing an organic cellular material along with at least two metallic, preferably steel balls such that a vertical motion of the sample box causes the provided balls to impact each other thereby disrupting the organic cells placed in each vial accommodated within the wells and releasing the cellular material.
A system for gravity assisted high throughput disruption of a plurality of organic cellular
materials comprising:
(a) a machine unit comprising a motor, a rotatable motor shaft connected to said motor and having one end protruding out of said motor, a second rotatable shaft substantially housed within-an apparatus casing and being rotatably connected to * the motor shaft through a coupling capable of transmitting rotatory motion from said motor shaft to the second shaft, a crank disposed on said second rotatable shaft which receives rotatory motion from the second shaft and converts the

received rotatory motion into reciprocating motion of a connecting rod connected thereto through a crank pin, a crank pin housed within said crank casing and disposed on said crank and adapted to accommodate a connecting rod, a connecting rod. being connected to said crank through said crank pin at a first end such that the reciprocating motion generated by the crank is received by the connecting rod through the crank pin and transmitted to a piston shaft connected at second end of said connecting rod through a provided second crank pin, a piston shaft having a first end connected to said connecting rod through the second crank pin and a second end protruding out of said apparatus casing, a platform placed on said protruding end of the piston shaft and adapted to hold a plurality of sample boxes, a clamping plate adapted to cover a plurality of sample boxes placed on the platform, said clamping plate having a plurality of apertures, each said aperture being threaded internally, a plurality of tie rods being externally threaded through a substantial length thereof such that said internal thread provided on said plurality of clamping plate apertures engages with said external thread provided on said plurality of tie rods to hold said sample boxes firmly in place; and a control unit comprising at least a depressive start and a. depressive stop button, a first display means displaying the number of oscillations per second conducted by the sample holder and adapted to allow an user of the system to digitally pre-set a predetermined oscillation frequency, a rotatable knob adapted to allow an user to vary the oscillation frequency during the operation of the machine unit, a second display means displaying the time elapsed since the initiation of operation of the machine unit and adapted to allow an user to digitally pre-set a predetermined operation time, an emergency stop button to terminate the operation in between, such that upon the start button being depressed by a user, the motor initiates a rotation in the motor shaft, coupling and the second shaft which is translated into a reciprocating motion of the connecting rod and the piston shaft, the oscillating piston shaft oscillates the sample box and the plurality of wells provided in said sample box, each said well accommodating a vial containing an organic cellular material along with at least two metallic, preferably steel balls such that a vertical oscillation of the sample box causes the provided

balls to impact each other thereby disrupting the organic cells contained in said organic cellular material placed in each vial accommodated within the wells.
A method for gravity assisted high throughput disruption of a plurality of organic
cellular material comprising:
(a) providing an apparatus comprising a motor, a rotatable motor shaft connected to said motor and having one end protruding out of said motor, a second rotatable shaft substantially housed within an apparatus casing and being rotatably connected to the motor shaft through a coupling capable of transmitting rotatory motion from said motor shaft to the second shaft, a crank disposed on said second rotatable shaft which receives rotatory motion from the second shaft and converts the received rotatory motion into reciprocating motion of a connecting rod connected thereto through a crank pin, a crank pin housed within said crank casing and disposed on said crank and adapted to accommodate a connecting rod, a connecting rod being connected to said crank through said crank pin at a first end such that the reciprocating motion generated by the crank is received by the connecting rod through the crank pin and transmitted to a piston shaft connected at second end of said connecting rod through a provided second crank pin, a piston shaft having a first end connected to said connecting rod through the second crank pin and a second end protruding out of said apparatus casing, a platform placed on said protruding end of the piston shaft and adapted to hold a plurality of sample boxes, a clamping plate adapted to cover a plurality of sample boxes placed on the platform, said clamping plate having a plurality of apertures, each said aperture being threaded internally, a plurality of tie rods being externally threaded through a substantial length thereof such that said internal thread provided on said plurality of clamping plate apertures engages with said external thread provided on said plurality of tie rods to hold said sample boxes firmly in place;
(b) providing a control unit for said apparatus, said control unit comprising at least a depressive start and a depressive stop button, a first display means displaying the number of oscillations per second conducted by the sample holder and adapted to

allow an user of the system to digitally pre-set a predetermined oscillation frequency, a rotatable knob adapted to allow an user to vary the oscillation frequency during the operation of the machine unit, a second display means capable of displaying the time elapsed since the initiation of operation of the machine unit and adapted to allow an user to digitally pre-set a predetermined operation time;
(c) selecting at least one sample box having a plurality of wells provided therein;
(d) selecting a plurality of sample vials and placing a plurality of cellular material samples in said sample vials;
(e) freezing the cellular material samples by placing said at least one sample box in liquid nitrogen;
(f) upon reaching a predetermined freezing temperature, placing said at least one sample box on said platform of provided apparatus and engaging said sample box between the platform and clamping plate firmly in place with a provided plurality of tie rods;
(g) initiating the provided motor for a predetermined amount of time which is less than about two minutes during which time the vertical oscillation of the sample box causes the provided balls to impact each other thereby disrupting the organic cells contained in said organic cellular material placed in each vial accommodated within the wells;
(h) optionally reinitiating the provided motor for another predetermined amount of time and allowing the motor to stop after the passage of a predetermined amount of time;
(i) adding an extraction buffer to the disrupted cellular material samples placed in said plurality of sample vials such that the disrupted samples are stabilized; and
(j) storing the stabilized cellular materials.
A method for gravity assisted high throughput extraction of a desired cellular material from a plurality of organic tissues comprising:
(a) providing an apparatus comprising a motor, a rotatable motor shaft connected to said motor and having one end protruding out of said motor, a second rotatable

shaft substantially housed within an apparatus casing and being rotatably connected to the motor shaft through a coupling capable of transmitting rotatory motion from said motor shaft to the second shaft, a crank disposed on said second rotatable shaft which receives rotatory motion from the second shaft and converts the received rotatory motion into reciprocating motion of a connecting rod connected thereto through a crank pin, a crank pin housed within said crank casing and disposed on said crank and adapted to accommodate a connecting rod, a connecting rod being connected to said crank through said crank pin at a first end such that the reciprocating motion generated by the crank is received by the connecting rod through the crank pin and transmitted to a piston shaft connected at second end of said connecting rod through a provided second crank pin, a piston shaft having a first end connected to said connecting rod through the second crank pin and a second end protruding out of said apparatus casing, a platform placed on said protruding end of the piston shaft and adapted to hold a plurality of sample boxes, a clamping plate adapted to cover a plurality of sample boxes placed on the sample plate, said clamping plate having a plurality of apertures, each said aperture being threaded internally, a plurality of tie rods being externally threaded through a substantial length thereof such that said internal thread provided on said plurality of clamping plate apertures engages with said external thread provided on said plurality of tie rods to hold said sample boxes firmly in place;
(b) providing a control unit for said apparatus, said control unit comprising at least a depressive start and a depressive stop button, a first display means displaying the number of oscillations per second conducted by-the sample holder and adapted to allow an user of the system to digitally pre-set a predetermined oscillation frequency, a rotatable knob adapted to allow an user to vary the oscillation frequency during the operation of the machine unit, a second display means displaying the time elapsed since the initiation of operation of the machine unit and adapted to allow an user to digitally pre-set a predetermined operation time;
(c) selecting at least one sample box having a plurality of wells provided therein:

(d) selecting a plurality of sample vials and placing a plurality of cellular material samples in said sample vials;
(e) freezing the cellular material samples by placing said at least one sample box in liquid nitrogen;
(f) upon reaching a predetermined freezing temperature, placing said at least one sample box on said platform of provided apparatus and engaging said sample box between the platform and clamping plate firmly in place with a provided plurality of tie rods;
(g) initiating the provided motor for a predetermined amount of time which is less than about two minutes during which time the vertical oscillation of the sample box causes the provided balls to impact each other thereby disrupting the organic cells contained in said organic cellular material placed in each vial accommodated within the wells;
(h) optionally reinitiating the provided motor for another predetermined amount of time and allowing the motor to stop after the passage of a predetermined amount of time;
(i) precipitating the cellular proteins and polysaccharides;
(j) centrifuging the precipitated proteins and polysaccharides;
(k) adding a binding buffer, in combination with ethanol, to cause the desired intracellular material to bind on a provided silica-based membrane; and
(1) eluting the desired cellular material in a low salt buffer or deionized water.
A kit-of-parts for gravity assisted high throughput disruption of a plurality of organic
cellular material comprising:
(a) a machine unit comprising a motor, a rotatable motor shaft connected to said motor and having one end protruding out of said motor, a second rotatable shaft substantially housed within an apparatus casing and being rotatably connected to the motor shaft through a coupling capable of transmitting rotatory motion from said motor shaft to the second shaft, a crank housed within said crank casing and disposed on said second rotatable shaft which receives rotatory motion from the second shaft and converts the received rotatory motion into reciprocating motion

of a connecting rod connected thereto through a crank pin, a crank pin housed within said crank casing and disposed on said crank and adapted to accommodate a connecting rod, a connecting rod being connected to said crank through said crank pin at a first end such that the reciprocating motion generated by the crank is received by the connecting rod through the crank pin and transmitted to a piston shaft connected at second end of said connecting rod through a provided second crank pin, a piston shaft having a first end connected to said connecting rod through the second crank pin and a second end protruding out of said apparatus casing, a platform placed on said protruding end of the piston shaft and adapted to hold a plurality of sample boxes, a clamping plate adapted to cover a plurality of sample boxes placed on the sample plate, said clamping plate having a plurality of apertures, each said aperture being threaded internally, a plurality of tie rods being externally threaded through a substantial length thereof such that said internal thread provided on said plurality of clamping plate apertures engages with said external thread provided on said plurality of tie rods to hold said sample boxes firmly in place; (b) a control unit comprising at least a depressive start and a depressive stop button, a first display means displaying the number of oscillations per second conducted by the sample holder and adapted to allow an user of the system to digitally pre-set a predetermined oscillation frequency, a rotatable knob adapted to allow an user to vary the oscillation frequency during the operation of the machine unit, a second display means displaying the time elapsed since the initiation of operation of the machine unit and adapted to allow an user to digitally pre-set a predetermined operation time;
(c) a plurality of sample boxes, each said sample box comprising a plurality of sample wells, each said sample well adapted to hold a sample vial;
(d) a plurality of metallic, preferably stainless steel balls having a diameter of about 1 mm to about 6 mm;
(e) a plurality of sample vials having a volume of about 1 ml to about 3 ml;
(f) a extraction reagent comprising detergent, antifoaming agent, buffering agent and ribonuclease A;

(g) a nitrogen bath with circulating liquid nitrogen for freezing the organic cellular
material placed in the sample vials; (h).a binding buffer comprising guanidine hydrochloride, diluted with about 96% to
about 100% ethanol; (i) a precipitation buffer comprising a predetermined concentration of acetic acid or a
predetermined concentration of ethanol; (j) a wash buffer comprising a predetermined concentration of 70% to 100% ethanol; (k) an elution buffer comprising a combination of a predetermined concentration of
Tris-Cl, ethylenediaminetetraacetic acid (EDTA), sterile distilled water or mixture
thereof; (1) a plurality of pipettes; and (m)an instruction manual comprising instructions for gravity assisted high throughput
disruption of a plurality of organic cellular material according to a predetermined
method.
The above description sets forth, rather broadly, a summary of one embodiment of the present invention so that the detailed description that follows may be better understood and contributions of the present invention to the art may be better appreciated. Some of the embodiments of the present invention may not include all of the features or characteristics listed in the above summary. There are, of course, additional features of the invention that will be described below and will form the subject matter of the present invention. In this respect, before explaining at least one preferred embodiment of the invention in detail, it is to be understood that the invention is not limited in its application to the details of the construction and to the arrangement of the components set forth in the following description or as illustrated in the drawings. The invention is capable of other embodiments and of being practiced and carried out in various ways. Also, it is to be understood that the phraseology and-terminology employed herein are for the purpose of description and should not be regarded as limiting.

BRIEF DESCRIPTION OF THE DRAWINGS
The invention will now be described in connection with the drawings described hereinafter.
Figure 1 represents a schematic representation of an apparatus for gravity assisted high throughput disruption of a plurality of organic cellular material according to the preferred embodiment of the present invention.
DETAILED DESCRIPTION OF THE INVENTION
Accordingly, in one aspect, the present invention provides an apparatus for gravity assisted high throughput disruption of a plurality of organic cellular material. The ' apparatus comprises a motor, a rotatable motor shaft, a second rotatable shaft, a crank, a crank pin, a connecting rod, a piston shaft, a platform, a clamping plate and a plurality of tie rods.
In an embodiment, the preferable motor is a motor having a power denomination of about 1 hp to about 2 hp, preferably about 1.5 hp.
In one embodiment, the motor shaft is preferably connected to a second rotatable shaft through a coupling. The coupling rotatably connects the protruding end of the motor shaft with a first end of second rotatable shaft.
A coupling is a known device conventionally used in the art to connect two shafts together at their ends for the purpose of transmitting power. However, the presence of a coupling is not particularly limiting and its need may be eliminated altogether.
Thus, in another non-limiting embodiment, the motor shaft and a second rotatable shaft may be combined into a single shaft which protrudes out of the provided motor and a substantial length of which resides within the apparatus body housed within the apparatus casing. In another embodiment, the protruding end of the motor shaft may be immovably connected to the first end of the second rotatable shaft.

The apparatus of the present invention comprises a crank disposed on the second rotatable shaft. The crank receives rotatory motion from the rotation of the second shaft and converts the received rotatory motion into reciprocating motion of a connecting rod connected thereto. Preferably, the connecting rod may be connected to the crank through a connecting crank pin.
In a preferred embodiment, a crank pin is provided on the crank and accommodates a connecting rod such that the rotatory motion of the crank is converted to a reciprocating vertical motion of the connecting rod.
In an embodiment, the provided crank and the crank pin are housed substantially within a crank housing.
In another embodiment, the reciprocating vertical motion of the connecting rod is transmitted to a connected piston shaft. The connecting rod is preferably connected to a piston shaft through a second crank pin. In an embodiment, the second crank pin is preferably disposed at the second end of the connecting rod where it connects the connecting rod with a piston shaft.
The piston shaft has a first end connected to the connecting rod through the crank pin. The second end of the piston shaft preferably protrudes out of the apparatus casing. In this embodiment, the apparatus casing comprises an aperture to allow protrusion of the piston shaft out of the apparatus casing.
In an embodiment, the piston preferably includes a bearing bush cylindrical lining provided onto said second end of the piston shaft to reduce the frictional contact between the frictional shaft and the aperture provided in the apparatus casing.
In another embodiment, the apparatus of the present invention comprises a supporting plate. The supporting plate supports the apparatus casing along with the components of the apparatus comprised therein including the second rotatable shaft, the crank casing, the crank, the crank pin, the connecting rod and the piston shaft etc. at the top face thereof.

The bottom face of the supporting plate comprises a plurality of cushion springs. The cushion springs absorb the vibrations generated during the operation of the apparatus and also serve to reduce the intensity of noise generated by the apparatus.
In another embodiment, the cushion springs are provided with a plurality of rubber bushes and help maintain the apparatus retained at a predetermined location. Preferably, the apparatus of the present invention comprises four cushion springs provided at four corners of the supporting plate. Each provided cushion spring further comprises a rubber bush such that the apparatus of the present invention comprises at least four rubber bushes. Each rubber bush, when firmly placed on the ground, substantially displaces air from its enclosed volume and creates a local vacuum at the chosen location which keeps the apparatus firmly placed at the chosen location despite the vibrations generated during the operation of the-apparatus.
The apparatus of the present invention comprises a platform provided at the protruding end of the piston shaft. The platform is adapted to hold a plurality of sample boxes, either placed adjacent or on top of each other. In a more preferred embodiment, the sample plate is substantially rectangular in shape.
In an embodiment, said platform is adapted to hold at least one sample box. More preferably, the platform is adapted to hold from two to about four sample boxes during one operation cycle of the apparatus. However, those skilled in the art will appreciate that the apparatus may be adapted to hold more than four sample boxes as well by increasing the dimensions of the platform although four sample boxes generally suffice.
In another embodiment, each sample box is capable of holding at least 96 samples. However, the number of wells provided in a sample box is not particularly limiting such that sample boxes containing upto 384 wells for holding 384 samples per sample box may also be conveniently used.

The apparatus further comprises a clamping plate, which covers the sample boxes placed on the platform. The clamping plate has a plurality of apertures, each of which is threaded internally. The apparatus includes a plurality of tie rods, which are each threaded externally.
. Preferably, the clamping plate comprises two apertures which accommodate two tie rods.
The internal thread provided on the clamping plate apertures engages with the external threads provided on the tie rods so as to hold the sample boxes firmly placed on the sample plate. A vertical reciprocating motion of the piston shaft causes a vertical motion of the sample box.
Each provided sample box has a plurality of wells provided therein. Preferably, each sample box includes 96 wells. In an embodiment, each provided well is adapted to accommodate at least one sample vial containing an organic cellular tissue. Each sample is further provided with at least two metallic balls.
In a preferred embodiment, the preferred metallic balls are stainless steel balls although it should be understood that other metallic balls are not excluded.
A vertical motion of the sample box causes the provided balls to impact each other thereby disrupting the organic cells contained in said organic tissue placed in each vial accommodated within the wells. It has been surprisingly found that the apparatus of the present invention is capable of simultaneously disrupting the organic cells of a large number of tissue samples i.e. preferably from about 24 samples to about 384 samples within about 30 seconds at a preferred oscillation frequency of about 800 to 1200 oscillations per minute at a stroke length of about 30 mm without causing an appreciable shear to the desired cellular material giving a maximum yield of the cellular material. The yield of the desired intracellular material was found to be greater than about 50 ng/μL.

The present inventors have found that extracted yield of the organic cellular material is maximum at an oscillation frequency of 800-1200 RPM at about 30 mm of stroke length within a time period of about 20 to 30 seconds. Trial runs were conducted using the apparatus of the present invention for extraction of DNA from tomato leaves. The following results were obtained when the quantification of DNA was done with UV Pharma Spec Visible Spectrophotometer (Shimadzu):
Table 1

SNo. Oscillation Stroke Time (s) Number Yield Qualitative
frequency (RPM) length (mm) of
operation
cycles (ng/μL) observations
1 300 30 20 10.25 Insufficient disruption
2 800 30 20 33.87 Good
3 1000 30 20 2 31.12 Good
4 1200 30 20 30.50 Good
5 1400 30 20 42.75 Highly sheared
6 1500 30 20 38.50 Highly sheared
7 1000 30 10 27.75 Insufficient disruption
8 1000 30 5 18.12 Insufficient disruption
9 1000 30 60 37.50 Highly sheared
10 1000 30 120 43.00 Highly sheared
The following conclusions were derivable from the experimental results listed above:

1. The maximum DNA yield was observed at 800-1200 RPM at 30 mm stroke length within about 20 to 30 seconds. At higher RPM measurements, the extracted DNA was highly sheared while at lower RPM measurements, the tissue disruption was insufficient for an appreciable yield.
2. The DNA yield decreased for time periods less than 20 seconds due to insufficient disruption or more than 30 seconds due to high shear of the extracted DNA.
3. The DNA yields are comparable for one or two disruption cycles.
Thus, in another aspect, the present invention provides a system for gravity assisted high throughput disruption of a plurality of organic cellular tissue. The system of the present invention comprises an apparatus for gravity assisted high throughput disruption of a plurality of organic cellular tissue.
In the embodiment, the system according to this aspect of the present invention comprises an apparatus described in any aspect described herein above relating to an apparatus for high throughput disruption of organic cellular tissue or any embodiment thereof, which are reproduced herein by reference in its entirety.
In addition, the system of the present invention comprises a control unit. In an embodiment, the control unit is provided attached to the apparatus for gravity assisted high throughput disruption of cellular tissue. In this embodiment, the control unit may be juxtaposed adjacent to the apparatus. In yet another embodiment, the components of the control unit are divided into at least two sub-units, which are disposed at each side of the apparatus. In this embodiment, the balance of the system according to the present invention, including the apparatus, is maintained in a stable position.
In another embodiment, the control unit is separately provided as an accompaniment to the apparatus of the present invention.

The control unit is adapted to control the operation of the apparatus according to the . present invention. The control unit comprises a depressive start and a depressive stop button which causes the apparatus to start or stop the operation on being manually depressed.
In the embodiment, the. control unit additionally comprises an emergency depressive stop button which immediately causes the apparatus to stop the operation at any time during its operation on being manually depressed.
The control unit comprises a first display means which is adapted to display the oscillation frequency conducted by the sample holder. In one embodiment, the display means displays the oscillation frequency measured per second or measured per minute. In one embodiment, the first display,means includes a pre-defined set of depressive buttons having individual digits marked therein so as to allow an user of the system to digitally pre-set a predetermined oscillation frequency.
The control unit comprises a rotatable knob, which upon rotation allows an user to regulate the oscillation frequency of the apparatus during its operation.
The control unit preferably comprises a second display means. The second display means displays the time elapsed since the initiation of the operation of the machine unit. In another embodiment, the second display means includes a pre-defined set of depressive buttons having individual digits marked therein so as to allow an user of the system to digitally pre-set a predetermined operating time for the apparatus of the present invention.
In this embodiment, the operation of the apparatus is initiated when the provided start button is manually depressed by an user of the apparatus, which causes the motor to initiate a rotation in the motor shaft, coupling and the second shaft which is translated into a reciprocating motion of the connecting rod and the piston shaft. The oscillating piston shaft oscillates the sample box and the plurality of wells provided in said sample box. Each well accommodates a vial containing an organic cellular tissue along with at least two balls such that a vertical oscillation of the sample box causes

the provided balls to impact each other thereby disrupting the organic cells contained in said organic tissue placed in each vial accommodated within the wells.
In another aspect, the present invention provides a method for gravity assisted high throughput disruption of a plurality of organic cellular material.
The method comprises providing an apparatus for disruption of the organic cellular tissue. In the embodiment, the method according to this aspect of the present invention comprises providing an apparatus described in any aspect described hereinabove relating to an apparatus for high throughput disruption of organic cellular tissue or any embodiment thereof, which are reproduced herein by reference in its entirety.
The method further comprises providing a control unit for controlling or regulating the operation of the provided apparatus. Thus, in a preferred embodiment, the method according to this aspect of the present invention comprises providing a control unit described in any aspect described hereinabove relating to a system for high throughput disruption of organic cellular tissue or any embodiment thereof, which is reproduced herein by reference in its entirety.
The method of the present invention comprises selecting at least one sample box having a plurality of wells provided therein. Preferably, a plurality of sample boxes may also be selected if the number of organic tissue samples exceeds the number of wells provided in one sample box. The method further comprises selecting a plurality of sample vials such that each sample vial accommodates one organic cellular tissue. The plurality of organic cellular tissue sample is placed into the plurality of sample vials.
In the embodiment, the sample vials are contacted with liquid nitrogen in a provided nitrogen bath to freeze the organic cellular tissue samples. In this embodiment, the sample vials are preferably brought to a temperature of at least about -196°C in order to ensure freezing of the organic tissue samples instantly.

At least one sample box containing the frozen tissue sample is placed on the platform of the apparatus. The sample box is firmly engaged between the sample plate and the clamping plate with at least one tie rod, preferably at least about four tie rods.
The apparatus motor is thereafter initiated with a pre-set operating time of at least about 20 seconds to about less than two minutes, preferably less than one minute. The vertical oscillation of the sample box along with the platform causes the provided balls to impact each other thereby disrupting the organic cells.
In another preferred embodiment, the apparatus motor is re-initiated with a pre-set operating time of at least about 20 seconds to about less than two minutes, preferably less than one minute. The apparatus motor is thereafter allowed to stop after the passage of the pre-set amount of time. The organic tissue sample is preferably recooled in a nitrogen bath being subjected to a second disrupting operation in the apparatus of the present invention.
The method of the present invention includes addition of an extraction buffer to the disrupted cellular material samples placed in said plurality of sample vials such that the intracellular material of the disrupted tissue sample is stabilized.
In an embodiment, the extraction buffer preferably comprises CTAB buffer alone or in combination with one or more reagents selected from p-mercapto-ethanol and isoamyl alcohol. Preferably, a 2% CTAB buffer may be used. CTAB (cetrimonium or hexadecyltrimethylammonium) buffer is a cationic surfactant which is a preferred buffer solution conventionally used for the extraction of nucleic material, particularly DNA.
The stabilized intracellular material may thereafter be stored for a desired period of time. In another embodiment, the stabilized intracellular material may be subjected to an extraction process for the extraction and/or isolation of the desired intracellular material.

Thus, in another aspect, the present invention provides a method for gravity assisted high throughput extraction of the intracellular material contained within an organic tissue. The method comprises disrupting the organic tissue according to the preceding aspect or any embodiment thereof.
Preferably, the intracellular material which is. extracted according to the present invention may be DNA or RNA though other intracellular material within a plant or an animal cell is not excluded. In an embodiment, other cellular materials such as proteins, alkaloids, plant hormones and polysaccharides may also be extracted using the apparatus and method of the present invention.
The method of the present invention comprises extracting the desired cellular material from the disrupted tissue sample using any conventional method known in the art.
In the embodiment, the cellular proteins and polysaccharides are precipitated using a salt known in the art. The precipitates and other unwanted cell debris are excluded by centrifugation. Thereafter, a known binding buffer, optionally in combination with ethanol, is added to promote the binding of the desired intracellular material such as DNA to a provided membrane. The sample is thereafter subjected to another centrifugation step which binds the cellular material to a provided membrane whereas contaminating cellular materials such as proteins and polysaccharides are removed by washing steps. The relatively pure desired cellular material is thereafter eluted in a small volume of a known low salt buffer or deionized water.
The said centrifugation steps may be carried out at about 6000 revolutions per minute for about 5 minutes in a provided centrifuge.
The preferred membrane, which selectively binds the intracellular material, is preferably a silica-based membrane, which selectively binds the desired DNA.
The preferred binding buffer used in this embodiment of the invention is a reagent comprising from about 96% to about 100% ethanol. The preferred eluting buffer

comprises a combination of a predetermined concentration of Tris-Cl and ethylenediaminetetraacetic acid (EDTA).
In an embodiment, the preferred stabilizing buffer and/or binding buffer for stabilizing the disrupted sample may be CTAB buffer, preferably a 2% strength solution thereof.
In another aspect, the present invention provides a kit-of-parts for gravity assisted high throughput disruption of a plurality of organic cellular material and extraction of a desired intracellular material therefrom.
The kit comprises a machine unit and a control unit. In this embodiment, the kit according to this aspect of the present invention comprises a machine unit including an apparatus described in any aspect described hereinabove relating to an apparatus for high throughput disruption of organic cellular tissue or any embodiment thereof, which are reproduced herein by reference in its entirety. The kit further comprises a control unit described in any aspect described hereinabove or any embodiment thereof, which are reproduced herein by reference in its entirety.
The kit comprises a plurality of sample boxes. Preferably, the kit comprises at least one sample box and upto four sample boxes. Each sample box comprises a plurality of sample wells, each sample well being adapted to hold a sample vial. The kit comprises a plurality of stainless steel balls having a diameter of about 1 mm to about 6 mm. Preferably, the stainless steel balls possess a diameter of from about 3 mm to about 4 mm.
In the embodiment, each sample vial included within the kit according to the present invention has a volume of about 1 ml to about 3 ml.
The kit further comprises a nitrogen bath with circulating liquid nitrogen for freezing the organic cellular material placed in the sample vial and for preventing the thawing and degradation of the desired intracellular material.

The kit according to the present invention comprises one or more buffers, which are tabulated as hereunder:

SNo. Buffer nomenclature Ingredients
1 Lysis buffer Detergent, Buffering agent, Ribonuclease A and antifoaming agent
2 Precipitation buffer Acetic acid and/or ethanol
3 Binding buffer Concentrated Guanidine hydrochloride, diluted with 100% ethanol.
4 Wash buffer Contains 70% to 100% ethanol
5 Elution buffer Tris-Cl + EDTA (optionally sterile distilled water may be used either alone or in combination with Tris-Cl and EDTA)'
The kit according to the present invention further comprises an instruction manual comprising instructions for cellular material extraction according to a predetermined method. Preferably, the instructions included within the instruction manual causes the method of the present invention recited in any preceding aspect or embodiment to be carried out.
Turning now to figure 1, illustrated is an apparatus for gravity assisted high throughput disruption of a plurality of organic cellular tissues. The apparatus comprises a motor (1) and a rotatable motor shaft (2) connected to the motor and having one end protruding out of said motor. A coupling (3) rotatably connects the protruding end of the motor shaft with a first end of a second rotatable shaft. A second rotatable shaft (4,5) is substantially housed within the apparatus casing (11) and is rotatably connected to the motor shaft through a coupling capable of transmitting rotatory motion from said motor shaft to the second shaft. A crank casing (6) substantially houses a crank and a crank pin. A crank housed within the crank casing and disposed on said second rotatable shaft receives rotatory motion from the second shaft and converts the received rotatory motion into reciprocating motion of a connecting rod connected thereto through a crank pin. A crank pin (7)

is housed within said crank casing and disposed on said crank and adapted to accommodate a connecting rod. A connecting rod (8) is connected to the crank through the crank pin at a first end such that the reciprocating motion generated by the crank is received by the connecting rod through the crank pin and transmitted to a piston shaft connected at second end of the connecting rod through a provided second crank pin. The second crank pin (7) is disposed at the second end of the connecting rod and connects the second end of the connecting rod with a piston shaft. A piston shaft (9) has a first end connected to the connecting rod through the second crank pin and a second end protrudes out of the apparatus casing (11). A.bearing bush (10) cylindrical lining is provided on the piston shaft to reduce the frictional contact between the piston shaft and an aperture (not shown) provided in the apparatus casing to allow protrusion of the piston shaft out of the apparatus casing. A supporting plate is adapted to support. at the top face thereof, the apparatus casing along with the second rotatable shaft, the crank casing, the crank, the crank pin, the connecting rod, the piston shaft along with the other sampling appurtenances. A plurality of cushion springs (12) is provided at the bottom face of the rectangular supporting plate. A plurality of. rubber bushes (13) is attached to the cushion springs and adapted to maintain the apparatus firmly placed at a predetermined location. A sample plate is placed on the protruding end of the piston shaft and adapted to hold a plurality of sample boxes. A clamping plate (14) is adapted to cover a plurality of sample boxes placed on the platform (16). The clamping plate has a plurality of apertures, each aperture being threaded internally. A plurality of tie rods (15) are externally threaded through a substantial length such that the internal thread provided on the plurality of clamping plate apertures engages with the external thread provided on the tie rod to hold the sample boxes firmly in place. A reciprocating motion of the piston shaft causes a vertical motion of the sample box. Each sample box has a plurality of wells provided therein. Each provided well accommodates a vial containing an organic cellular material along with at least two balls such that a vertical motion of the sample box causes the provided balls to impact each other

thereby disrupting the organic cells contained in said organic cellular tissue placed in each vial accommodated within the wells.
Experiment 1
Tomato leaves were used to extract DNA using the method of the present invention as the protocol for extraction. About 40.0 mg of tomato leaves were taken for each trial experiment and free dried in liquid nitrogen. The free dried tomato leaves were powdered in the apparatus of the present invention. DNA was extracted from the disrupted material using the methods described herein. The conclusions from the gel photographs corresponded with the conclusions derivable from the results tabulated above.
Wherein the aforegoing reference has been made to integers or components having known equivalents, then such equivalents are herein incorporated as if individually set forth. Accordingly, it will be appreciated that changes may be made to the above described embodiments of the invention without departing . from the principles taught herein. Additional advantages of the present invention will become apparent for those skilled in the art after considering the principles in particular form as discussed and illustrated. Thus, it will be understood that the invention is not limited to the particular embodiments described or illustrated, but is intended to cover all alterations or modifications which are within the scope of the present invention.

WE CLAIM
1. An apparatus for gravity assisted high throughput disruption of a plurality of organic cellular material comprising:
(a) a motor;
(b) a rotatable motor shaft connected to said motor and having one end protruding out of said motor;
(c) a second rotatable shaft substantially housed within an apparatus casing and being rotatably connected to the motor shaft;
(d) a crank disposed on said second rotatable shaft which receives rotatory motion from the second shaft and converts the received rotatory motion into reciprocating motion of a connecting rod connected thereto through a crank pin;
(e) a crank pin disposed on said crank and adapted to accommodate a connecting rod;
(f) a connecting rod being connected to said crank through said crank pin at a first end such that the reciprocating motion generated by the crank is received by the connecting rod through the crank pin and transmitted to a piston shaft connected at second end of said connecting rod through a provided second crank pin;
(g) a piston shaft having a first end connected to said connecting rod through the second crank pin and a second end protruding out of said apparatus casing;
(h) a platform placed on said protruding end of the piston shaft and adapted to
hold a plurality of sample boxes; (i) a clamping plate adapted to cover a plurality of sample boxes placed on
the platform, said clamping plate having a plurality of apertures, each said
aperture being threaded internally; and (j) a plurality of tie rods being externally threaded through a substantial
length thereof such that said internal thread provided on said plurality of
clamping plate apertures engages with said external thread provided on

said tie rods to hold said sample boxes firmly in place such that a reciprocating motion of said piston shaft causes a vertical motion of the sample box, each said sample box having a plurality of wells provided therein, each said well accommodating a vial containing an organic cellular material along with at least two metallic, preferably steel balls such that a vertical motion of the sample box causes the provided balls to impact each other thereby disrupting the organic cells placed in each vial accommodated within the wells and releasing the cellular material.
2. The apparatus as claimed in claim 1, wherein said motor has a power denomination of about 1 hp to about 2 hp, preferably about 1.5 hp.
3. The apparatus as claimed in claim 1 or claim 2, wherein protruding end of said motor shaft is connected to first end of said second rotatable shaft through a coupling.
4. The apparatus as claimed in any preceding claim additionally comprising a supporting plate capable of supporting the apparatus casing along with the second rotatable shaft, the crank casing, the crank, the crank pin, the connecting rod and the piston shaft.
5. The apparatus as claimed in claim 4, wherein the bottom face of said supporting plate comprises a plurality of cushion springs adapted to absorb the vibrations generated during the operation of the apparatus.
6. A system for gravity assisted high throughput disruption of a plurality of organic cellular materials comprising:
(a) a machine unit comprising a motor, a rotatable motor shaft connected to said motor and having one end protruding out of said motor, a second rotatable shaft substantially housed within an apparatus casing and being rotatably connected to the motor shaft through a coupling capable of transmitting rotatory motion from said motor shaft to the second shaft, a crank disposed on said second rotatable shaft which receives rotatory motion from the second shaft and converts the

received rotatory motion into reciprocating motion of a connecting rod connected thereto through a crank pin, a crank pin housed within said crank casing and disposed on said crank and adapted to accommodate a connecting rod, a connecting rod being connected to said crank through said crank pin at a first end such that the reciprocating motion generated by the crank is received by the connecting rod through the crank pin and transmitted to a piston shaft connected at second end of said connecting rod through a provided second crank pin, a piston shaft having a first end connected to said connecting rod through the second crank pin and a second end protruding out of said apparatus casing, a platform placed on said protruding end of the piston shaft and adapted to hold a plurality of sample boxes, a clamping plate adapted to cover a plurality of sample boxes placed on the platform, said clamping plate having a plurality of apertures, each said aperture being threaded internally, a plurality of tie rods being externally threaded through a substantial length thereof such that said internal thread provided on said plurality of clamping plate apertures engages with said external thread provided on said plurality of tie rods to hold said sample boxes firmly in place; and (b) a control unit comprising at least a depressive start and a depressive stop button, a first display means displaying the number of oscillations per second conducted by the sample holder and adapted to allow an user of the system to digitally pre-set a predetermined oscillation frequency, a rotatable knob adapted to allow an user to vary the oscillation frequency during the operation of the machine unit, a second display means displaying the time elapsed since the initiation of operation of the machine unit and adapted to allow an user to digitally pre-set a predetermined operation time, an emergency stop button to terminate the operation in between, such that upon the start button being depressed by a user, the motor initiates a rotation in the motor shaft, coupling and the second shaft which is translated into a reciprocating motion of the connecting rod and the piston shaft, the oscillating piston shaft oscillates the sample box and the plurality of wells provided in said sample box, each said well accommodating a vial containing an organic cellular material along with at least two metallic, preferably steel balls such that a vertical

oscillation of the sample box causes the provided balls to impact each other thereby disrupting the organic cells contained in said organic cellular material placed in each vial accommodated within the wells.
7. The system as claimed in claim 6, wherein said control unit is separately provided as an accompaniment or alternately, being attached to the apparatus such as. herein described.
8. A method for gravity assisted high throughput disruption of a plurality of organic cellular material comprising:
(a) providing an apparatus comprising a motor, a rotatable motor shaft connected to said motor and having one end protruding out of said motor, a second rotatable shaft substantially housed within an apparatus casing and being rotatably connected to the motor shaft through a coupling capable of transmitting rotatory motion from said motor shaft to the second shaft, a crank disposed on said second rotatable shaft which receives rotatory motion from the second shaft and converts the received rotatory motion into reciprocating motion of a connecting rod connected thereto through a crank pin. a crank pin housed within said crank casing and disposed on said crank and adapted to accommodate a connecting rod, a connecting rod being connected to said crank through said crank pin at a first end such that the reciprocating motion generated by the crank is received by the connecting rod through the crank pin and transmitted to a piston shaft connected at second end of said connecting rod through a provided second crank pin, a piston shaft having a first end connected to said connecting rod through the second crank pin and a second end protruding out of said apparatus casing, a platform placed on said protruding end of the piston shaft and adapted to hold a plurality of sample boxes, a clamping plate adapted to cover a plurality of sample boxes placed on the platform, said clamping plate having a plurality of apertures, each said aperture being threaded internally, a plurality of tie rods being externally threaded through a substantial length thereof such that said internal thread provided on said plurality of clamping plate apertures engages with said

external thread provided on said plurality of tie rods to hold said sample boxes firmly in place;
(b) providing a control unit for said apparatus, said control unit comprising at least a depressive start and a depressive stop button, a first display means displaying the number of oscillations per second conducted by the sample holder and adapted to allow an user of the system to digitally pre-set a predetermined oscillation frequency, a rotatable knob adapted to allow an user to vary the oscillation frequency during the operation of the machine unit, a second display means capable of displaying the time elapsed since the initiation of operation of the machine unit and adapted to allow an user to digitally pre-set a predetermined operation time;
(c) selecting at least one sample box having a plurality of wells provided therein;
(d) selecting a plurality of sample vials and placing a plurality of cellular material samples in said sample vials;
(e) freezing the cellular material samples by placing said at least one sample box in liquid nitrogen;
(f) upon reaching a predetermined freezing temperature, placing said at least one sample box on said platform of provided apparatus and engaging said sample box between the platform and clamping plate firmly in place with a provided plurality of tie rods;
(g) initiating the provided motor for a predetermined amount of time which is less than about two minutes during which time the vertical oscillation of the sample box causes the provided balls to impact each other thereby disrupting the organic cells contained in said organic cellular material placed in each vial accommodated within the wells;
(h) optionally reinitiating the provided motor for another predetermined amount of time and allowing the motor to stop after the passage of a predetermined amount of time; and
(i) extracting the desired cellular material from said disrupted cellular material.

9. The method as claimed in claim 8, wherein said apparatus is operated at an oscillation frequency of about 800 to 1200 revolutions per minute.
10. The method as claimed in claim 8 or claim 9, wherein said motor is initiated for about 20 seconds to about 30 seconds.
11. The method as claimed in claims 8-10 wherein said apparatus is operated at a stroke length of 30 mm.
12. The method as claimed in claims 8-11, wherein extracting the desired cellular material from said disrupted cellular material comprises (a) adding an extraction buffer to the disrupted cellular material samples placed in said plurality of sample vials such that the disrupted samples are stabilized; and (b) storing the stabilized cellular materials.
13. The method as claimed in claims 8-11, wherein extracting the desired cellular material from said disrupted cellular material comprises (a) precipitating the cellular proteins and polysaccharides; (b) centrifuging the precipitated proteins and polysaccharides; (c) adding a binding buffer, in combination with ethanol, to cause the desired intracellular material to bind on a provided silica-based membrane; and (d) eluting the desired cellular material in a low salt buffer or deionized water.
14. A kit-of-parts for gravity assisted high throughput disruption of a plurality of organic cellular material comprising:
(a) a machine unit comprising a motor, a rotatable motor shaft connected to said motor and having one end protruding out of said motor, a second rotatable shaft substantially housed within an apparatus casing and being rotatably connected to the motor shaft through a coupling capable of transmitting rotatory motion from said motor shaft to the second shaft, a crank housed within said crank casing and disposed on said second rotatable shaft which receives rotatory motion from the second shaft and converts the received rotatory motion into

reciprocating motion of a connecting rod connected thereto through a crank pin, a crank pin housed within said crank casing and disposed on said crank and adapted to accommodate a connecting rod, a connecting rod being connected to said crank through said crank pin at a first end such that the reciprocating motion generated by the crank is received by the connecting rod through the crank pin and transmitted to a piston shaft connected at second end of said connecting rod through a provided second crank pin, a piston shaft having a first end connected to said connecting rod through the second crank pin and a second end protruding out of said apparatus casing, a platform placed on said protruding end of the piston shaft and adapted to hold a plurality of sample boxes, a clamping plate adapted to cover a plurality of sample boxes placed on the sample plate, said clamping plate having a plurality of apertures, each said aperture being threaded internally, a plurality of tie rods being externally threaded through a substantial length thereof such that said internal thread provided on said plurality of clamping plate apertures engages with said external thread provided on said plurality of tie rods to hold said sample boxes firmly in place;
(b) a control unit comprising at least a depressive start and a depressive stop button, a first display means displaying the number of oscillations per second conducted by the sample holder and adapted to allow an user of the system to digitally preset a predetermined oscillation frequency, a rotatable knob adapted to allow an user to vary the oscillation frequency during the operation of the machine unit a -second display means displaying the time elapsed since the initiation of operation of the machine unit and adapted to allow an user to digitally pre-set a predetermined operation time;
(c) a plurality of sample boxes, each said sample box comprising a plurality of sample wells, each said sample well adapted to hold a sample vial;

(d) a plurality of metallic, preferably stainless steel balls having a diameter of about 1 mm to about 6 mm;
(e) a plurality of sample vials having a volume of about 1 ml to about 3 ml;
(f) a extraction reagent comprising detergent, antifoaming agent, buffering agent and ribdnuclease A;
(g) a nitrogen bath with circulating liquid nitrogen for freezing the organic cellular material placed in the sample vials;
(h) a binding buffer comprising guanidine hydrochloride, diluted with
about 96% to about 100% ethanol; (i) a precipitation buffer comprising a predetermined concentration of
acetic acid or a predetermined concentration of ethanol; (j) a wash buffer comprising a predetermined concentration of 70% to
100% ethanol; (k) an elution buffer comprising a combination of a predetermined
concentration of Tris-Cl, ethylenediaminetetraacetic acid (EDTA),
sterile distilled water or mixture thereof; (1) a plurality of pipettes; and (m)an instruction manual comprising instructions for gravity assisted high
throughput disruption of a plurality of organic cellular material
according to a predetermined method.
15. The kit-of-parts as claimed in claim 14, wherein one or more buffers are selected from a lysis buffer, a precipitation buffer, binding buffer, wash buffer and elution buffer wherein constituents of said one or more buffers are selected according to the below table:

SNo. Buffer nomenclature Ingredients
1 Lysis buffer Detergent, Buffering agent, Ribonuclease A and antifoaming agent
2 Precipitation buffer Acetic acid and/or ethanol
3 Binding buffer Concentrated Guanidine hydrochloride, diluted with 100% ethanol.
4 Wash buffer Contains 70% to 100% ethanol
5 Elution buffer Tris-Cl + EDTA (optionally sterile distilled water may be used either alone or in combination with Tris-Cl and EDTA)
16. The kit-of-parts as claimed in claim 14 or claim 15, wherein the instructions included within the instruction manual causes the method such as herein described to be carried out.
17. An apparatus for gravity assisted high throughput disruption of a plurality of organic cellular material substantially as described herein with reference to the example and accompanying drawing.
18. A system for gravity assisted high throughput disruption of a plurality of organic cellular, material substantially as described herein with reference to the example and accompanying drawing.
19. A method for gravity assisted high throughput disruption of a plurality of organic cellular material substantially as described herein with reference to the example and accompanying drawing.

20. A kit-of-parts for gravity assisted high throughput disruption of a plurality of organic cellular material substantially as described herein with reference to the example and accompanying drawing.