WO2001059159A2

WO2001059159A2 - Molecular biological kit apparatus and method

Info

Publication number: WO2001059159A2
Application number: PCT/US2001/004028
Authority: WO
Inventors: Jack Chirikjian
Original assignee: INSTASTAIN
Current assignee: INSTASTAIN
Priority date: 2000-02-10
Filing date: 2001-02-09
Publication date: 2001-08-16
Anticipated expiration: 2002-08-10
Also published as: WO2001059159A3; AU2001238065A1

Abstract

The invention provides an apparatus that may be used as a kit for biological experiments. The apparatus includes a multi-well container, a first biological reagent in one well of the container, a second, different biological reagent in a second well of the container, and a penetrable film covering the container and sealing the contents of the plate. The penetrable film includes printing on the film identifying the contents of the wells. The biological reagents may be in solid or liquid form in the wells, and may include enzymes, substrates, polymeric beads and buffers. The invention may be used as a kit for research or teaching purposes, and may be prepared as a customized kit by a manufacturer.

Description

TITLE

MOLECULAR BIOLOGICAL KIT APPARATUS AND METHOD

CROSS REFERENCE TO RELATED APPLICATIONS AND CLAIM FOR BENEFIT OF

EARLIER FILING DATE

This application claims all benefits under 35 U.S .C .1 1 9(e) from my United States Provisional Application entitled MOLECULAR BIOLOGICAL KIT APPARA TUS AND METHOD, serial number 60/1 81 ,560, filed February 10, 2000.

BACKGROUND OF THE INVENTION

Field of the Invention

The present invention relates to the field of molecular biology, particularly to methods and apparatus for molecular biological assays, and particularly to the packaging of molecular biological kits.

Description of the Related Art

The use of molecular biology kits has greatly enhanced the efficiency of performing biological research. Here, "molecular biology" is used to encompass a number of areas of biology in which small volume assays may be performed, including those generally associated with molecular biology and nucleic acids, as well as cellular biology, protein chemistry, and immunology. A kit provides the important reagents necessary for performing a particular protocol, greatly simplifying the preparation for performing the protocol and providing a consistency and repeatability which is difficult to achieve otherwise.

For example, a conventional kit for a restriction enzyme digest might contain a tube of an enzyme such as EcoR I , a tube of 1 0X reaction buffer concentrate, and perhaps a tube of gel loading buffer concentrate. Enzyme samples are typically provided in 1 .5-mL tubes with snap or screw caps, and must be stored in the freezer. Buffers are provided in separate tubes that are often stored separately from the enzymes by the user. A typical kit might be useful for digestion of 20 samples, for example. Although such a kit is extremely useful, with each use the tubes of the kit must be located, withdrawn from storage, opened, appropriately aliquoted to follow the particular protocol, and then stored again for future use of the remaining reagent. To insure consistency, the various reagents from the same lot should be used together, even if stored separately. Sometimes, repeated freezing and thawing of the samples can induce loss of activity of the components.

As an improvement on the providing of enzymes to be stored in the freezer, some enzymes are available commercially in freeze-dried, pre-aliquoted form, but these are still typically provided in small snap-cap tubes, separately from buffers and other reagents that might be used with the enzymes.

Kits are useful for teaching as well as research purposes, but when conducting a teaching laboratory in molecular biology, the problems of using a conventional kit are exacerbated. Because a number of students will be performing a particular protocol at different locations in the laboratory, each student must have samples of each of the reagents to be used. For reasons of cost, small amounts of enzyme and reagents must be aliquoted for each student. Buffer solutions must also be prepared and aliquoted. This is time-consuming and tedious for the teacher or laboratory manager, and an error in preparation will ruin the exercise for the students. Moreover, students are often still learning basic skills of laboratory work, and they may have difficulty labeling, tracking and manipulating the tubes, and in following written protocols and record keeping.

In the conventional art, there have been attempts to simplify molecular biological assays. Examples of the conventional art are seen, for example, in the following U.S. Patents.

U.S. Patent 4,925,629, to Schramm, entitled DIAGNOSTIC DEVICE, describes a device in the form of a kit for an immunoassay. The device has a housing including a microtiter plate, standard tubes for analyte standards, a buffer container and a container for radioactive or enzyme-labeled constituent. The device also may contain antibody adhered to the wells of the microtiter plate. The wells of the microtiter plate are removable.

U.S. Patent 5,073,341 , to Hargreaves, entitled DEVICES FOR CONDUCTING SPECIFIC BINDING ASSA YS, describes devices that may be made in multiwell plates with a penetrable septum that may be foil, polyethylene or rubber. The patent describes methods for forming a self-contained reaction mixture upon addition of an analyte. The patent also describes the use of cushions in the devices or hollow caps to serve as reagent reservoirs for additional reagent components.

U.S. Patent 5,252,774, to Honig et al., entitled REAGENT RECEPTACLE AND SUPPORT RACK FOR AUTOMA TED CLINICAL ANAL YZERS, describes a trapezoidal receptacle for a clinical analyzer, with a membrane spanning the top of the receptacle.

U.S. Patent 5,840,573, to Fields, entitled MOLECULAR ANAL YZER AND METHOD OF USE, describes an apparatus having a frame with an internal chamber for sealingly holding a number of sample tubes containing test samples and having pre-incorporated internally contained detection reagents. These reagents are injected between cavities in the device.

U.S. Patent 5,604, 101 , to Hanley et al. , entitled METHOD OF MINIMIZING CONTAMINA TION IN AMPLIFICA TION REACTIONS USING A REACTION TUBE WITH A PENETRABLE MEMBRANE, and U.S. Patent 5,753, 1 86, entitled REACTION TUBE WITH A PENETRABLE MEMBRANE TO MINIMIZE CONTAMINA TION, describe a disposable reaction vessel containing all of the reagents necessary for performing a nucleic acid amplification. The vessel is an individual tube with a membrane cover.

U.S. Patent 5,780,248, to Milchanoski et al., entitled FOIL SEALED CASSETTE FOR AGGLUTINA TION REA CTIONS AND LINER THEREFOR, describes a multi-well cassette with a barrier for retaining reactants in an upper chamber separated from a lower chamber.

However, the above-described apparatus and methods do not provide a convenient way to deal with the above-described problems of research and teaching kits for manually conducted assays. Based on my reading of the art, then, I have decided that what is needed is an improved method of packaging kits for biological protocols.

SUMMARY OF THE INVENTION

Therefore, it is an object of the present invention to provide an improved packaging arrangement and method of packaging of a molecular biological kit.

It is a further object of the invention to provide a packaging arrangement of a molecular biological kit that avoids the necessity of storing and manipulating individual reagent tubes.

It is a yet further object of the present invention to provide a molecular biology test kit that does not waste reagent.

A still further object of the invention is to provide a test kit that simplifies the performance of certain assays.

Another object of the invention is to provide an improved molecular biological kit for experiments in a teaching laboratory.

Yet another object of the invention is to provide a biological kit for a teaching laboratory that minimizes preparation work by the instructor.

Still another object of the invention is to provide a kit that simplifies the performance of an experiment by the student. These and other objects are achieved in the present invention, which provides an apparatus for biological experimentation including a multi-well container, a first biological reagent in one well of the container, a second, different biological reagent in a second well of the container, and a penetrable cover covering the container and sealing the contents of the plate. The penetrable cover includes a printed label identifying the contents of the wells. The biological reagents may be in solid or liquid form in the wells, and may include enzymes, substrates, polymeric beads, microencapsulated reagents, buffers, bacteria and yeast. The penetrable cover may be a film made of a material such as plastic or aluminum, and is generally penetrable by a pipet tip.

The multi-well container may be a multi-well plate, such as a 96-well plate, or may be a strip of tubes. In one embodiment, the multi-well plate is scored to allow individual rows of the plate, each having biological reagents, to be snapped off from the plate. Alternatively, a thin plate may be cut with scissors to separate rows.

The apparatus of the present invention may be used as a biological kit for research or teaching purposes. When used as a teaching kit, the apparatus may be organized to allow each separated row of the apparatus to serve as a kit for a single experiment for a student or groups of students. Alternatively, one apparatus may contain the reagents for a number of different experiments to be performed by a student.

BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1 is a plan view of one embodiment of an apparatus of the present invention; and

Fig. 2 is a cross-sectional view through line ll-ll of Fig. 1 . DETAILED DESCRIPTION OF THE INVENTION

The present invention involves an assay kit to be used in biological assays. Here, "biological" is taken to include the fields of microbiology, cellular biology, molecular biology, etc. The apparatus and methods of the present invention are applicable to any reactions or assays generally performed in small tubes or wells of plates. Turning now to the drawings, an embodiment of an assay kit according to the present invention is seen in Figure 1 .

The apparatus of the present invention generally includes a multi-well container. The embodiment shown in Figure 1 illustrates the features of an exemplary kit for performing a teaching laboratory experiment in which a 96-well plate 1 70 is the multi-well container. The 96-well plate 1 70 has rows that are easily separable. Although the rows shown in Fig. 1 are oriented in the short direction, that is, rows of 8 wells, the rows could alternatively be oriented in the long direction. Score marks 1 50 allow the rows to be snapped off by hand. The wells of the 96-well plate are covered with penetrable cover 1 60, which is a film for sealing reagents in the well and that can be penetrated by a disposable pipet tip, such as the type of tip commonly used with a pipettor to transfer reagents. Typical materials for the penetrable cover include plastic and aluminum.

In the example of Fig. 1 , penetrable cover 1 60 is printed with labels 1 10, 1 20, and 1 30 on one of the snap-off rows. For clarity, only the first row is shown with printing in Figure 1 and only some labels are shown, but generally each well to be used would be labeled. In this example, the printing on the penetrable film includes label 1 20 identifying the particular apparatus or the particular snap-off row of the apparatus, label 1 1 0 which is a number identifying a particular well, and label 130 which is text identifying the contents of a particular well. The wells 1 40 of the 96-well plate, shown dashed because they lie below the penetrable film, are filled with appropriate reagents, and in one embodiment, each row of the plate would contain the same reagents as each other row, thus allowing 1 2 identical rows to be snapped off by the laboratory teacher and distributed to the students. As shown in Fig. 2, which is a cross section of one of the rows of Fig. 1 , the wells of a row of plate 1 70 contain reagents sealed in the wells by the penetrable cover 1 60. In the example of Fig. 2, three of the wells contain reagents. Well 21 0 contains an aliquot of one liquid reagent, and well 220 contains an aliquot of another liquid reagent. Well 230 contains a lyophilized reagent in powder form. It is also possible to have a well with a reagent adsorbed onto the well, or a well containing a microencapsulated reagent or containing polymer beads conjugated to biological reagents. The other wells, including well 240, in this example are empty. Wells of the plate may be empty either because fewer than one row's worth of reagents are necessary for the kit, or because these wells are reserved for use as reaction vessels during use of the kit. The wells shown in Fig. 2 are round- bottomed by way of example. However, wells of the present invention may be flat- bottomed, micro-test tube-shaped, conical, or of other shapes known in the art.

In another embodiment, a multi-well plate that can be cut into strips of wells may be used rather than a snap-off plate. Here, once sealed, the multi-well plate may be cut along lines indicated on the sealing film. If the plate has vertical edges along the border of the plate, the cut strip may be freestanding. If not, a holder can be provided to hold the strip so that the wells remain vertical while in use.

The embodiment illustrated in Fig. 1 uses a 96-well plate. However, the invention is readily applied to other multi-well plates, such as 384-well plates. In another embodiment, a strip of tubes is used instead of a multi-well plate. In keeping with the invention, the strip is sealed with a penetrable film. In this embodiment, individual tubes may be separated from the strip for steps such as incubation.

When used for research purposes, a researcher will generally know the contents of each well of the plate. For example, if obtained commercially, an instruction sheet may accompany to plate to identify the contents. When used for teaching purposes, each removable row of the plate, for distribution to a different student, could be slightly different, containing, for example, different unknown samples in well 1 , but the same reagents in the other rows, in this case, the label 120 would be unique for each row to allow the laboratory teacher to know the contents of the unknown well.

In another embodiment, each row of a plate for a student laboratory could contain reagents for a separate experiment. In this embodiment, one student could, for example, be issued one plate for an entire laboratory course, and could snap-off or cut off the appropriate row at the beginning of a laboratory class.

The apparatus of the present invention when used for teaching will generally contain most of the reagents required for the student to perform an experiment. The included reagents may even include water. The apparatus may include empty wells for performing reactions. Thus, in the use of a teaching apparatus of the present invention, the student may aliquot reagents from the apparatus into reaction tubes, or may aliquot reagents into empty wells provided in the apparatus. The contents of one well may be used in a single reaction, or the well may contain sufficient contents for aliquoting to several tubes or wells, for several reactions.

The following Examples illustrate some of the possible applications to which the device of the present invention could be applied.

Example 1

Example 1 is a kit for a teaching laboratory experiment for "introduction to electrophoresis". Generally, such a kit will have samples of DNA of various molecular sizes, which can be used as markers or for simply learning how to perform electrophoresis.

A particular example would be a 96-well plate with 1 2 separable rows of 8 wells each. Each group of 8 wells would have 8 DNA samples of different molecular sizes, one sample in each well. The samples would be provided in liquid or lyophilized form. The samples could be provided with loading buffer already present, to simplify preparation by the student. In use, each student would receive one separated row of wells. The student would puncture each well, remove the sample, and load the sample onto a gel for DNA electrophoresis, and then electrophorese the samples.

Example 2

Example 2 is for a "restriction enzyme digest" experiment of a teaching laboratory. Generally, such a plate would contain between 1 and 6 different restriction enzymes, between 1 and 6 DNA samples, such as bacteriophage or viral DNAs, reaction buffers, distilled water and digestion reaction stop solution that includes a gel tracking dye.

In a particular example, a 96-well plate could be organized into eight separable rows of 1 2 wells each. The exemplary contents of one row of this teaching apparatus of the present invention are as follows: Column 1 , that is, the first well in each row, would contain a sample of unknown DNA. Column 2 would contain a lyophilized restriction enzyme in sufficient quantity for a single reaction, for example EcoRI . Columns 3 and 4 would likewise contain aliquots of two other restriction enzymes, for example BamH I and Hind3. Column 5 would contain a sample of λ-DNA, and columns 6, 7 and 8 would again contain lyophilized aliquots of EcoRI , BamH I and Hind3. Column 9 would contain a restriction enzyme reaction buffer, and column 10 would contain a DNA gel loading buffer concentrate. To use the kit, the student would puncture the well in column 9, containing buffer, and would aliquot this buffer to wells 1 and 5 to dissolve the DNA samples. The student would then aliquot the sample from well 1 to wells 2, 3 and 4, and from well 5 to wells 6, 7, and 8, and incubate the strip for about half an hour. This may be conveniently performed in a heating block. If it is necessary to cap the tubes during this time, a rubber pad can be compressed over the strip.

Then, the loading buffer concentrate would be aliquoted to wells 1 , 2, 3, 6, 7 and 8. The contents of these wells would then be ready to be loaded on a gel and electrophoresed. Thus, with a minimal number of pipetting steps and a high probability of success, the student would have performed the restriction digest. This particular example illustrates the provision of wells for performing reactions, obviating separate tubes for performing reactions.

Example 3

This example is a multi-plate well container that is a "restriction enzyme digestion plate" which contains from 1 to 6 different restriction enzymes, from 1 to 6 viral or bacteriophage DNAs, reaction buffers, distilled water, and digestion reaction stop solution including gel tracking dye.

Alternatively, a series of predigested DNAs in gel loading buffer can be provided, without providing restriction enzymes. With this plate, a student can simply load a gel with the contents of the wells.

Example 4

This example is a "protein analysis plate" for a student laboratory. Each separable row of the plate contains several native proteins, SDS/mercaptoethanol- denatured proteins, buffers and reagents to denature proteins.

A specific example would be a plate with 1 2 rows of eight wells each. The eight wells would contain three different proteins in native form, three denatured proteins, gel loading solution and denaturing solution.

Example 5

This example is a multi-well plate container that is an "immunology analysis plate: which contains a number of arrays of antigens such as proteins, primary antigens, secondary antigens which are enzyme-linked and one of more color substrates that are typically used for ELISA and other immunological reactions. A specific example would be a plate with 1 2 rows of eight wells each. Two different antigens, two related primary antibodies, the two related secondary antibodies, and the two different substrates would be provided in the eight wells.

Example 6

This example is kit that is a "cloning and PCR analysis plate" that contains the biologicals and reagents for cloning and transformation and the reagents for polymerase chain reaction (PCR) and reverse-transcriptase polymerase chain reaction (RT-PCR) . The reagents for cloning and transformation include several restriction enzymes, DNA ligase, and plasmids. The PCR reagents include Taq DNA polymerase, dATP, dGTP, dCTP and dTTP, a sample of the DNA of interest and the oligonucleotide primers for PCR. Here, "oligonucleotide" will generally refer to single stranded nucleic acid molecules of relatively short length, typically less than about 100 nucleotides. As such, "oligonucleotides" will be distinguished from larger pieces of DNA or RNA which might serve, for example, as templates. For RT-PCR, a few RNA templates and primers and reverse transcriptase will be included. It should be possible to provide all of the reagents necessary for a complete cloning, PCR and RT-PCR on a single plate, except for host cells for cloning.

Example 7

A variation on Example 6 is a "DNA profiling (fingerprinting) plate which would contain different variable number of tandem repeat (VNTR) primers, mitochondrial primers, Taq DNA polymerase, a mixture of the four dNTPs, a standard DNA ladder and a gel loading solution.

Example 8

This example is a "molecular biology dye plate" that can be used to demonstrate various electrophoresis experiments that simulate DNA fragment migration. The dye patterns can be used to simulate experiments in molecular biology such as DNA sizing, mapping and DNA fingerprinting. Typically, the multi- well container will be separable into rows of eight wells each, providing well dyes or mixture of dyes for electrophoresis. For example, five to ten different organic dyes selected for their mobilities could be provided either as premixed solutions or separately. Gel loading solution would typically also be provided in one well.

Example 9

This example encompasses a number of possible kits for reactions of nucleic acids, which may be used for molecular biological research. The plates could be organized in various ways, including, for example, one detachable row for each sample to be processed. Sample kits would contain aliquoted reagents appropriate for reactions such as DNA ligation, PCR, RT-PCR, restriction digestion. Other possible kits include aliquoted markers for protein RNA and DNA analysis.

For example, a plate for standard restriction enzyme digests could be made in a 96-well plate, and could have eight rows of 1 2 wells. The 1 2 wells would each have a unit aliquot of a restriction enzyme, forming a restriction enzyme panel. Alternatively, 1 2 rows of eight wells each are possible, as well as other combinations with other sized plates. Alternatively, these panels could be provided in strips of tubes, rather than in a separable plate.

Example 10

This example is a "bacterial/yeast sample plate". This plate would contain a series of bacteria and/or yeast strains in separate wells of a detachable row of a plate. These bacterial/yeast samples may be provided frozen or dried. A typical preparation would involve aliquoting a suspension of a bacterial or yeast strain in an appropriate medium into each well. The medium may include a cryogenic or lyophilization stabilizer, for example trehulose or dry milk, as appropriate. The samples would then be lyophilized and sealed, or sealed then frozen. This plate can be used for microbiology research or teaching laboratories. For example, the plate could be used in fermentation experiments. Advantages of such a plate over conventional bacterial preparation include lowered cost, miniaturization, reduced waste disposal, containment, reduced time and more reliable preparation.

Example 1 1

In this Example, which is a variation on Example 6, one well of a multi-well plate would contain Taq DNA polymerase, dATP, dCTP, dGTP, TTP and appropriate reaction buffer. Another well of the plate would contain at least one pair of oligonucleotide primers. To use this embodiment of the invention, one would combine the contents of the first and second wells, and add a sample of DNA to be analyzed. This would lead to a sample solution ready for polymerase chain reaction analysis.

Example 1 2

In this Example, wells of a multi-well container would contain a series of different buffers or other related reagents to be compared. In one embodiment, a series of buffers differing in pH could be provided in different wells. In another embodiment, a series of buffers differing in salt concentration could be provided in different wells. In another embodiment, a series of enzyme inhibitors could be provided in different wells. This arrangement would allow comparison of enzyme activity under different conditions. Embodiments of this Example might be used for the purpose of optimizing reaction conditions for an enzyme or for testing an enzyme against a panel of inhibitors. Alternatively, embodiments of this Example might be used to provide a convenient source of a variety of buffers, such as restriction enzyme buffers.

Example 1 3

In this Example, at least one well of the plate contains a polymer bead or lyophilized pellet which contains a biological reagent. Such a bead or pellet may have several different chemicals present, including chemicals which might be incompatible if stored in the same solution. For example, polymer beads are known in the art which contain dATP, dCTP, dGTP and dTTP, Taq DNA polymerase, reaction buffer and a stabilizer. This plate could be used for DNA amplification studies as in previous Examples.

Kits made using the apparatus and method of the present invention could be of use to research scientists in a variety of fields, as well as for clinical, forensic, home diagnostic or other routinely performed assays. In teaching, the apparatus of the present invention would be useful for teaching laboratories of courses from middle school to graduate level. In addition, the kits could be useful for the amateur scientist market, home "biology sets" for youngsters, etc.

A method of making a molecular biological kit according to the present invention will now be described . First, a multi-well container is provided . As noted for the apparatus embodiments above, this container may be a connected strip of tubes or a plate such as a 96-well plate. In the case of using a plate with a matrix arrangement of wells, a plate with snap-off rows or columns may be used.

Then, any reagents that are to be provided as lyophilized powders in the final kit are introduced into the wells dissolved in an appropriate carrier solvent. For example, to provide a dehydrated buffer, an aqueous solution of that buffer would be added. Enzymes to be provided would be introduced in an appropriate buffer. Certain reagents of low water solubility may be introduced in volatile organic solvents. An appropriate volume of the reagent solution is added to deliver the appropriate amount of solute for the well .

Then, if lyophilized powders are to be provided, the plate is lyophilized by an appropriate technique. These techniques may include vacuum concentration, centrifugal vacuum concentration, freeze-drying or air-drying.

After the lyophilization step, reagents to be found in liquid form in the finished device are introduced to the appropriate wells. Then, the plate is sealed with the penetrable cover or film. The sealing process may be performed using heat-sealing or using adhesive, by methods well known in the art. Generally, the penetrable film will already be printed with the desired information labeling the wells However, in some circumstances it may be desirable to print some information on the film after the sealing process. This may include lot information or indicia.

Generally, the biological test kits of the present invention will be made to be stable at room temperature. In some cases, however, the entire plate may be refrigerated or frozen as necessary.

From a commercial standpoint, the present invention is readily applicable to a customized kit that could be manufactured and shipped to the user. Here, the user would specify the reagents to be supplied in the multi-well plate, and would communicate an order to the manufacturer The manufacturer would then prepare a customized plate by aliquoting the appropriate reagents as described above, printing a customized penetrable film, and sealing the plate.

For example, a customizable plate might be a restriction digest panel, such as described in Example 9, above However, with a customizable plate, the user would be able to select from a large list of available enzymes. There are plainly many permutations of available enzymes, and a customizable arrangement would allow for any permutation to be available to the customer without the need for the manufacturer to stock a separate kit for each permutation.

The examples given here serve only to illustrate the various embodiments of the invention. It will be readily apparent to those skilled in the art that the structures, shapes, materials and other parameters may be modified or substituted without departing from the scope and spirit of the invention.

Claims

WHAT IS CLAIMED IS:

1 . A biological apparatus, comprising: a multi-well container; an aliquot of a first biological reagent in a first well of said multi-well container, said first biological reagent not comprising an oligonucleotide; an aliquot of a second biological reagent, different from said first biological reagent, in a second well of the container; a penetrable cover covering the wells of the multi-well container; and printing on the outer surface of said penetrable cover, for identifying the contents of said first and second wells.

2. The biological apparatus of claim 1 , wherein said penetrable cover is made of metal foil or plastic.

3. The biological apparatus of claim 1 , wherein said aliquot of a first biological reagent is liquid at room temperature.

4. The biological apparatus of claim 1 , wherein said aliquot of a first biological reagent is a lyophilized solid.

5. The biological apparatus of claim 1 , wherein said first well further comprises a polymer bead containing the first biological reagent.

6. The biological apparatus of claim 1 , wherein said first biological reagent comprises a protein.

7. The biological apparatus of claim 1 , wherein said first biological reagent comprises a first double-strand DNA of greater than 200 base pairs; and wherein second biological reagent comprises a second double-stranded DNA which is different from the first double-stranded DNA.

8. The biological apparatus of claim 7, wherein said first bilogical reagent further comprises an additional double-stranded DNA of different molecular sizes, for use as markers.

9. The biological apparatus of claim 8, further comprising an aliquot of a third bilogical reagent in a third well of the multi-well container, said third biological reagent comprising a third double-stranded DNA which is different from said second double-stranded DNA.

10. The biological apparatus of claim 6, said protein comprises an enzyme.

1 1 . The biological apparatus of claim 6, wherein said enzyme is a restriction enzyme.

1 2. The biological apparatus of claim 1 , wherein said first biological reagent is a buffer.

1 3. The biological apparatus of claim 1 2, wherein said buffer comprises a gel loading buffer.

14. The biological apparatus of claim 1 , wherein said multi-well container is a microtube strip.

1 5. The biological apparatus of claim 1 , wherein said multi-well container is a multi-well plate.

1 6. The biological apparatus of claim 1 5, wherein said multi-well container is a 96-well plate.

1 7. The biological apparatus of claim 1 5 , wherein said multi-well container is a 384-well plate.

18. The biological apparatus of claim 1 5 , wherein each row of said multi-well plate comprises: aliquots of said first and second biological reagents, the arrangement of the aliquots of the first and second biological reagents in each row being the same as in the other rows of the plate.

1 9. The biological apparatus of claim 1 7, wherein each row of the plate comprises: lyophilized aliquots of at least three different restriction enzymes respectively in separate wells of the row.

20. The biological apparatus of claim 1 9, wherein each row of the plate further comprises an aliquot of a restriction enzyme reaction buffer in another well of the row.

21 . The biological apparatus of claim 20, wherein each row of the plate further comprises an aliquot of DNA gel loading buffer in another well of the row.

22. The biological apparatus of claim 1 , wherein said first biological reagent comprises a strain of bacteria or yeast.

23. The biological apparatus of claim 1 , wherein: said first biological reagent comprises Taq polymerase; said second biological reagent comprises dATP, dGTP, dCTP and dTTP; and further comprises: an aliquot of a third biological reagent comprising an oligonucleotide primer in a third well of the multi-well container.

24. The biological apparatus of claim 1 , wherein: said first biological reagent comprises Taq polymerase, dATP, dCTP, dGTP and dTTP; and said second biological reagent comprises two oligonucleotide primers.

25. The biological apparatus of claim 24, further comprising: an aliquot of a third reagent in a third well of the multi-well container, said third reagent comprising two oligonucleotide primers, said third reagent being different from said second reagent.

26. The biological apparatus of claim 1 , wherein said first biological reagent is a first enzyme reaction buffer and wherein said second biological reagent is a second enzyme reaction buffer.

27. The biological apparatus of claim 26, wherein said first enzyme reaction buffer has a different pH value than said second enzyme reaction buffer.

28. The biological apparatus of claim 26, wherein said first enzyme buffer differs from said second biological buffer in a concentration of a salt.

29. The biological apparatus of claim 1 , wherein said first biological reagent is a first enzyme inhibitor and said second biological reagent is a second enzyme inhibitor.

30. A biological apparatus, comprising: a multi-well plate having scoring between rows of the plate for snapping off rows of the plate; an aliquot of a first biological reagent in a first well of said multi-well plate; an aliquot of a second biological reagent, different from said first biological reagent, in a second well of the multi-well plate; a penetrable cover covering the wells of the multi-well plate; and printing on the outer surface of said penetrable cover, for identifying the contents of said first and second wells.

31 . The biological apparatus of claim 30, wherein said first biological reagent comprises a first double-strand DNA of greater than 200 base pairs; and wherein second biological reagent comprises a second double-stranded DNA which is different from the first double-stranded DNA.

32. The biological apparatus of claim 31 , wherein said first bilogical reagent further comprises an additional double-stranded DNA of different molecular sizes, for use as markers.

33. The biological apparatus of claim 32, further comprising an aliquot of a third bilogical reagent in a third well of the multi-well plate, said third biological reagent comprising a third double-stranded DNA which is different from said second double- stranded DNA.

34. A method of teaching biology, comprising the steps of: (i) providing a biological apparatus, said apparatus comprising a multi-well container, an aliquot of a first biological reagent in a first well of said multi-well container, said first biological reagent not comprising an oligonucleotide, an aliquot of a second biological reagent, different from said first biological reagent, in a second well of the multi-well container, a penetrable cover covering the wells of the multi-well container, and a label printed on the outer surface of said penetrable cover, for identifying the contents of said first and second wells;

(ii) puncturing the penetrable cover on said first well of the multi-well container with a pipet tip; and

(iii) dispensing solution from the pipette tip into said first well of the multi-well plate into the pipet tip.

35. The method of claim 34, further comprising: after the step of dispensing solution, withdrawing solution from the first well; puncturing the penetrable cover on said second well of the multi-well container with a tip of the pipet; and dispensing the solution from the pipet tip into the second well of the multi-well container.

36. The method of claim 34, , further comprising the steps of : before puncturing the penetrable cover on said first well, separating a row of the multi-well container from the remainder of the multi-well container, said row containing said first and second well; and performing the steps of puncturing the penetrable cover on the first well and dispensing solution into the first well, on the separated row of the multi- well plate.

37. The method of claim 34, wherein said first biological reagent is a protein.

38. The method of claim 36, further comprising the step of: separating additional rows of the multi-well plate, said multi-well plate having an aliquot of said first molecular biological reagent in one column position of each row of the multi-well container, and having an aliquot of said second molecular biological reagent in another column position of each row of the multi-well container.

39. A method comprising the steps of: providing a biological apparatus, said apparatus comprising: a multi-well container; an aliquot of a first biological reagent in a first well of said multi-well container; an aliquot of a second biological reagent, different from said first biological reagent, in a second well of the plate; a penetrable cover covering the wells of the multi-well container; and a label printed on the outer surface of said penetrable cover, for identifying the contents of said first and second wells; puncturing the penetrable cover on said first well of the multi-well container with a pipet tip; withdrawing solution in said first well of the multi-well container into the pipet tip; puncturing the penetrable cover on said second well of the multi-well container with the pipet tip; and dispensing the solution from the pipet tip into the second well of the multi- well container.

40. The method of claim 39, wherein said first biological reagent comprises an enzyme.

41 . A method of preparing a molecular biological apparatus, comprising the steps of: aliquoting a first molecular biological reagent to a first well of a multi-well plate, said first molecular biological reagent not comprising an oligonucleotide; aliquoting a second molecular biological reagent to a second well of a multi- well plate; printing a label on a penetrable cover for overlaying the plate near positions corresponding to said first and second wells, said label for identifying the first and second wells; and sealing said penetrable cover onto the multi-well plate.

42. The method of claim 41 , wherein said first biological reagent is an enzyme.

43. The method of claim 41 , wherein said step of aliquoting the first biological reagent comprising placing in the well a polymer bead containing the first biological reagent.