CN101233224A

CN101233224A - Recovery system of DNA and RNA or protein fragments with agarose gel or polyacrylamide gel

Info

Publication number: CN101233224A
Application number: CNA2006800280275A
Authority: CN
Inventors: 郑在景
Original assignee: Individual
Current assignee: Individual
Priority date: 2005-08-01
Filing date: 2006-08-01
Publication date: 2008-07-30
Also published as: KR20070118567A

Abstract

This is a device that directly collects DNA, RNA, or protein on agarose gel or polyacrylamide gel, which is separated and purified after applying electrophoresis to DNA, RNA, or protein on agarose gel or polyacrylamide gel. The present invention has improved the traditional methods that collect DNA, RNA, or protein fragments in the gel by slicing the gel in order to collect DNA, RNA, or protein fragments that are purified and separated by applying electrophoresis to DNA, RNA, or protein. The present invention offers a system that after applying electrophoresis to agarose gel or polyacrylamide gel, confirms the electrophoresed DNA, RNA, or protein fragments, and then collects DNA by sending DNA, RNA, or protein to the desired location through another elec not trophoresis, where the system has the integrated process of extracting DNA, RNA, or protein depending on the direction of the flow of electric charge or the reversed direction, and extracts DNA, RNA, or protein by utilizing the electrophoresis system different from the traditional one.

Description

From sepharose or polyacrylamide gel, reclaim the system of DNA, RNA or protein fragments

Technical field

Biotechnology, molecular biotechnology, biotechnology platform, electrochemistry, electroanalytical chemistry.

The invention relates to a kind of being used for from the technology of sepharose or polyacrylamide gel DNA isolation, RNA or protein fragments.The present invention has introduced after using sepharose or polyacrylamide gel (hereinafter will be called gel) electrophoresis, a kind of method in the whole bag of tricks of dna fragmentation, RNA fragment or the protein fragments that exists in the collection gel.

Under the situation of sepharose, electrophoresis is mainly used in DNA or RNA, and under the situation of polyacrylamide gel, electrophoresis is mainly used in protein, but can be used for DNA or RNA once in a while.

Background technology

After using electrophoresis, usually the DNA in the gel, RNA or protein fragments are cut into gel film.In order to improve required segmental selectivity, they should be cut into slices along every swimming band.There is several method to can be used for from the gel of section, collecting DNA, RNA or protein.

Method commonly used is that the gel of section is put into dialysis tubing, fills suitable buffered soln, reuses electrophoresis then in electrophoresis chamber.So the fragment in the gel will occur, the other guide thing in the collection tube except that gel.Can collect required fragment by separating collected buffered soln and fragment.

Perhaps, behind the gel that uses the section of pharmaceutical chemicals or heating for dissolving, directly put into micro glass beads, magnetic substance or resin each fragment is combined.By granulated glass sphere, magnetic substance or the thin resin of separation and combination, can therefrom collect required fragment then.

Compare protein, more different methods can be used for collecting DNA or RNA fragment.Film is put into syringe cylinder, and the sepharose of will cutting into slices is then put into wherein.Pressure pushing syringe piston with suitable grinds gel downwards.Promote piston always, stick to DNA on the film with collection.

On the gel of section, drip or two buffered soln.Gel is placed between negative ray and the anodic ray, electric charge is sent to buffered soln and gel by power supply.So the DNA in the gel will be pushed in the buffered soln.By this way, can extract DNA contained in the buffered soln.

On two sides of plastics tubing with cover, form the hole and put film.The gel of section is put into pipe, and after closing the lid, film is placed to gel and aligns.Pipe is put into electrophoresis chamber, make electric charge be sent to the place of on two sides, placing film, so that can extract DNA, RNA or protein in the gel.After the certain hour, in the time will extracting DNA, RNA in the gel or protein, the reversing pipe is so that make it move on to the reverse direction of flow of charge before extracting DNA, RNA or protein from film.Buffered soln in the take-off pipe is also collected fragment wherein then.

Can use various technology as mentioned above, but the most general technology is to use Filter column.A filter paper that will contain glass element is put into post.Gel is cut into slices, use the pharmaceutical chemicals dissolving, make dissolved solution penetrate into Filter column then.Fragment is combined, so that can extract DNA by washing with glass element in the filter paper.It is a kind of that to extract proteinic popular approach be only to use the method for resin.This method is a kind of method of the most generally using, and about the concrete technology of this special methods just in rapid progress, it will become the important core in the bio-science industry.

Summary of the invention

Technical problem

The ultimate aim of bio-science industry is the extension of quality life.In order to reach this target, each field and form are all in development.Wherein, in the presence of the task of many easy imaginations, the molecular biology paces of advancing are very slow.Research from the DNA field to the protein field comprises the most basic DNA, a little high-grade RNA and near the proteinic discovery of purpose, in succession with simultaneously underway.

These research projects are made of different fragmentary works usually.They take different approaches usually, but have only minority can form the combination of finishing short section plan when requiring.These work majorities are finished with hand, and need to consider well-educated people from job specification.For those reasons, the pace of progress of research is inevitably than desired see slow of scientist.For scientist, research steps is the same with the speed maintenance of scientist's brain soon will be very desirable.This needs require a kind of technology that is called as automatization, and automatization mainly is used for the analysis of base sequence in DNA research.Yet the stage before the base sequence analysis still is made of fragmentary works.

Because the problems referred to above, technology of the present invention proposes a kind of technical scheme of automatization, and for save time, the technology clue of high reproducibility and following automatization paves the way.

Technical scheme

In the present invention, behind use sepharose and the polyacrylamide gel electrophoresis, will be positioned on the gel with less spacing with the film (11) of DNA, RNA or protein fragments same widths.On film, form slit (12) and insert buffered soln or electrolytic solution, connect iontophoretic electrode (21) (+,-) then on the top of buffered soln or electrolytic solution.Although often connect positive electrode, the segmental electric charge according to be collected can connect negative potential.Accompanying drawing has shown how to handle the electrophoresis chamber lid (Fig. 2, Fig. 3) that is connected with electrode (21).The electrode (Fig. 2) that is connected with electrophoresis chamber lid (Fig. 2, Fig. 3) is immersed in buffered soln or the electrolytic solution by slit.In case finish electrophoresis on gel (14), the power supply (31,32) by blocking direction of fragments and it is turned to and cover the electrode (21) (Fig. 2 b) that is connected then makes DNA, RNA or protein fragments flow to buffered soln or electrolytic solution in the slit (12).Collect DNA, RNA or protein fragments by assembling buffered soln or electrolytic solution then.

Bottom (14) in slit (12) is porose, and bottom herein refers to the top of the sepharose that contacts with slit or the one side (93) of polyacrylamide gel.Slit (12) constitutes (Fig. 7) by a slice film, and extraction DNA, RNA or protein need be more than the faces (Fig. 7) of a picture film.Supply with buffered soln or electrolytic solution to the film another side, and on buffered soln of supplying with or electrolytic solution, apply electrophoresis power (62).

Top or the inside placement slit of gel or the one side of film at gel, for the slit that constitutes by slit, slit or more than one slit, be limited in the slit by beginning or end, DNA, RNA or protein fragments can be gathered in the slit electric charge.The difference of this method that Here it is and traditional method, traditional method are only with by collecting gel slice DNA, RNA or protein fragments.

There are several known methods to be used for collecting protein, DNA and polyacrylamide from sepharose.The something in common of these methods is must be with gel slice behind electrophoresis, and stops electrophoresis for section.

If study eight experiments carrying out at same target in succession, just be readily appreciated that structure of the present invention.Using common electrophoresis to carry out DNA, RNA or proteinic separation and confirm providing following experiment condition under the situation of process.Following condition is based on the situation of level.The same terms also can be used for vertical situation, in the case, the position transferred to vertically by level can obtain identical result.

The first, when using sepharose or polyacrylamide gel electrophoresis (Fig. 5), fragment flows to positive electrode (42) from negative potential (41) usually.For gel slab (43), form hole (44) and use buffered soln filling orifice (44) in direction of fragments.When using electrophoresis, various fragments (45) are in case arrive the hole arrives the hole with regard to passing (48) hole rapidly another side.Make it luminous even fluorescent substance or dyestuff are used for fragment, but still can not observe directly the scene that fragment is passed (48) hole with bore hole.At this moment, what can observe (48) is that the fragment (47) that centers on the hole disappears from negative potential, occurs in positive electrode direction then.If the buffered soln in chopping current and the collection hole (49) can extract very small amount of various fragment at this moment.

If can absorb the material of DNA, RNA or protein fragments, promptly micro glass beads, magnetic substance or resin are put into buffered soln or electrolytic solution, take out together then, and efficient can greatly improve so.

Second, when using sepharose or polyacrylamide gel electrophoresis, if between power cathode (51) and positive source (52), non-conducting material and gel (53) are coupled together, will form with the non-conductor same widths can not electrophoretic shade (54) section.If but with allowing the free-pouring conductor of electric current (53) to replace non-conductor to be connected with it, then from conductor to the negative potential direction electrophoresis (57) can take place.If use electrophoresis to arrive positive source, then from the conductor to the positive source, can form shade (57) until fragment.

The 3rd, according to above-mentioned second experiment combined conductor and non-conductor, form film (65) in this way, make conductor (62) be connected, and non-conductor (61) is connected with negative potential with positive electrode.Conductor (63) is configured to the top (66) of non-conductor (65) overlapping, and covers a certain height to non-conductor top.It is called " two film " popular name as it.After using gel electrophoresis, connect " two film " (Fig. 7, Fig. 8) to waiting to collect segmental positive electrode direction, that is, and the positive dirction of fragment swimming belt edge.When " two film " when being connected with gel slab, conductor should be faced positive electrode direction, and non-conductor should be faced the negative potential direction, and gel should not contact the conductor (62) on the non-conductor top of " two film ".The non-conductor direction on electrolytic solution or buffered soln edge " two film " is splashed into, i.e. the negative potential direction.At this moment, do not allow the diffusion of electrolytic solution or buffered soln, be with but make it rest on swimming to be extracted.Flow of charge is from positive electrode, then by the conductor of " two film ", conductor (62) and the electrolytic solution or the buffered soln on " two film " non-conductor top, until arriving fragment.Fragment was advanced along the reverse direction of flow of charge before being stoped by electrolytic solution or buffered soln.In addition, if use transfer pipet to pipette buffered soln, then can collect various fragments simply.Therefore, use this experiment can collect fragment on the gel.

The 4th, can use another kind of experimental technique to make " two film " by different way.Add the film of making by non-conductor (73) to the one side of " the two film " that contact with buffered soln or electrolytic solution, make buffered soln or electrolytic solution indiffusion and be retained in (Figure 10, Figure 11) in the safety zone." two film " of Zhi Zuoing is also referred to as " two membrane pores " by this way, but not the space between conductor thin film and " the two film " is called as " slit ", is also sometimes referred to as " hole ".When electrophoresis finishes, adjust " slit " to the top (74) of waiting to collect fragment swimming band." two membrane pores " (Figure 10, Figure 11) is connected with gel, before reusing electrophoresis, buffered soln or electrolytic solution put into " hole " then.By this way, can from the electrolytic solution of " slit " or buffered soln, collect DNA or RNA fragment.

If " two film " bottom of " two membrane pores " is cut rather than be connected, then can obtain the result shown in (Figure 12) with gel.Only remove the conductor part of positive electrode and directly positive source is connected with its top.Block electrophoretic positive source and provide electric current to the top of the positive source that is connected with electrophoretic power cathode and " two membrane pores ".So can collect DNA, RNA or protein fragments and buffered soln or electrolytic solution from " slit ".

The 5th, as shown in figure 15, making has " two membrane pores " of different conductor position.Build a bridge (101) in the bottom, and on bridge, form a dividing plate (101) as curtain." two membrane pores " (Figure 16) put into will be on the swimming lane of the isolating sample of electrophoresis.

When the fragment of confirming to extract in will gel is just in time below slit, stop ongoing electrophoresis, power to the power supply of conductor connection portion and relevant power supply.So from negative potential (111) direction, promptly any x axle (113) direction, the fragment that flows to positive electrode (112) direction will move to any y axle (114) and any z axle (115) direction simultaneously.Y axle (114) refers to the vertical direction of x axle (113) horizontal plane herein, and z axle (115) refers to the vertical direction of x axle and y axis plane.The fragment that moves will be included in the buffered soln or electrolytic solution in slit or hole.

This is in the situation of " two membrane pores ", and the axial bottom of y (116) is more downward than the axial bottom of x (117).Therefore, when using many swimming lanes electrophoresis, can stop each fragment of neighbouring lane to be mixed mutually.The height that should be higher than " two membrane pores " y axle bottom from the height (92) at the bottom of the gel at the bottom of the gel pore.

The 6th, above-mentioned experimental result shows that DNA, RNA or protein fragments can obtain part by electrophoresis and collect.When using many swimming lanes electrophoresis, must outfit can collect how segmental device is with in swimming.Therefore, should be with their " two membrane pores " level and vertical placement one by one.Directly connect the conductor and the positive source of membrane pores, perhaps remove conductor fully, and the positive electrode pin directly is immersed in buffered soln or the electrolytic solution from " two membrane pores ".When making sepharose or polyacrylamide gel, the position in the hole in the gel (94) should with membrane pores (promptly, slit) Dui Qi x axle (95) direction, and the bottom surface height (94) of the hole in the gel (94) (that is the basal surface position of the hole in the gel (92)) should be higher than the height of y axle (102) the direction bottom of " two membrane pores " bottom.The accidental blended possibility of the fragment of neighbouring lane when this method will be avoided extracting fragment.By buffered soln or electrolytic solution are put into slit, and use the positive current of slit and the negative current of same axis to extract fragment.

The 7th, in the time of after using electrophoresis, will collecting fragment, with " two membrane pores " in an overlapping form (Figure 17) place on sepharose (Figure 13) or the polyacrylamide gel (93)." two membrane pores " should have conductor in the positive electrode the inside.Can make " two membrane pores " according to the state of Figure 12, wherein cut the bottom that it is used to connect, rather than shown in the 4th experiment, be connected with gel.

After on any x axis, using electrophoresis, keep negative current and close positive current.Use the conductor-powered of " two membrane pores ", and be electrophoresis supply negative current, be the conductor supply positive current of " two membrane pores ".When being used for after slit is assembled the fragment required time and finished, cutting off the electricity supply, and collect buffered soln in the slit or electrolytic solution to collect fragment wherein.

Last method is from making agar or the polyacrylamide gel plate shown in (Figure 18).As Figure 19 and shown in Figure 20, " two membrane pores " is made of conductor (131) and non-conductor (132).When desiring to be placed on when making agar or polyacrylamide be converted into gel on the gel slab, be placed on behind the gel slice on the solidified gel slab.Use non-conductor as slit.When " two membrane pores " when placing gel with curing gel, guaranteed that solate can not flow between slit.After fragment that check to use electrophoresis to carry out is separated and is used buffered soln or electrolytic solution to fill slit, make electric charge from slit (132) effluent to fragment (133) side, deliver to slit (132) side with DNA, RNA or protein fragments (133) that will be to be collected.By this way, DNA, RNA or protein fragments (133) are collected in the slit (134).

Under the electrophoretic situation of the many swimming lanes of needs, a plurality of pins are placed between the swimming lane, use electrophoresis then as mentioned above in turn.

Beneficial effect

Foregoing invention had been integrated in the past and had been confirmed the independent extraction work that (next step of conventional electrophoretic work) back is done at RNA, DNA or protein fragments, had therefore saved the time and allowed to replace uncorrelated multistage manual operation by non-stop run.Present method has also added the new process of quantification and purifying in having only the electrophoresis process that separates and confirm function.

Electrophoretic technique can't be brought into play its repertoire, collect electrolytic material because also have no idea, and no matter whether it is the part of electrolytic action, but electrophoretic technique has been started a new era, promptly directly extracts DNA, RNA even protein from tissue, cell, animal blood and plant by promoting to collect.

This will be a basic technology that makes the manual operation automatization, particularly improve the reproducibility of work, and will become a useful invention of bio-science field.

Description of drawings

Fig. 1 has shown gel slab of the present invention.

Fig. 2 has shown to gel slab provides power supply.

Fig. 3 has shown the rear section that the electrode of power supply is provided to gel slab, and wherein the circuit at the back side and electrode are separated.

Fig. 4 has shown gel slab of the present invention.

Fig. 5 has shown that fragment is passed the order in hole when porose the existence.

Fig. 6 has shown conductor and idioelectric arrangement when using electrophoresis.

Fig. 7 and Fig. 8 have shown basic form and its principle of operation of fragment collection device among the present invention.

Fig. 9, Figure 10 and Figure 11 have shown than Fig. 7 and Fig. 8 and have more effectively collected segmental device.

Figure 12 has shown than Fig. 9, Figure 10 and Figure 11 and has more effectively collected segmental device.

Figure 13 and Figure 14 have explained distance and the x direction of principal axis between gel surface and the hole.

Figure 15 shown when collecting a large amount of fragment simultaneously, not the principle that can be polluted by neighbouring lane.

Figure 16 has briefly shown the device of collecting a large amount of fragments simultaneously and not polluted by neighbouring lane.

Figure 17 has shown no matter how the fragment size collects a large amount of segmental devices simultaneously from same sample.

Figure 18, Figure 19 and Figure 20 have shown the equipment that helps to produce a large amount of wieldy collection devices.

The description of reference numerals of accompanying drawing major portion

11: film 12: slit 13: gel foundry goods and gel slab 14: gel

16: hole, to be purified and putting into wherein of collecting such as experimental subjectss such as DNA, RNA, protein and tissues

31,32,41,42,51,52,71,72,81,111,112: the electrode of power supply or lead

44: the hole of containing buffered soln or electrolytic solution

45:DNA, RNA or protein fragments

53: non-conductor

55: conductor

62,63,66: the plate that comprises conductor

65: the thin plate of making by non-conductor

53: non-conductor

55: conductor

62,63,66: the plate that comprises conductor

65: the thin plate of making by non-conductor

74: the membrane pores that comprises segment conductor and idioelectric dual plate

92: place the hole of experimental subjects and the thickness between the gel outer wall

103: about in order not polluted, and should be thinner than the explanation of thickness 92 by neighbouring lane

113,114,115: about the detailed theoretical explanation of x, y and z axle electrophoresis part

Embodiment

In order to make maximum effect of the present invention, this will be the best mode of separating gel from collection device, and after separation by carrying out affirmation process and collection process independently, the user uses the new technology explanation and need not to break away from existing method and just can obtain facility.

The invention embodiment

As the gel foundry goods of placing gel among Fig. 1, the cover of Fig. 2 a is as downside, and Fig. 2 b is as upside.Their combination makes it possible to separate, purifying and collection.

Industrial applicibility

The experiment that great majority carry out in the biological gene engineering laboratory normally needs a large amount of manually operated phase I experiments. The present invention will have very great help to these experiments, and by as the technical background that makes the manual operation automation, will make substantial contribution for the life science process.

Statement according to the 19th modification of Patent Cooperation Treaty

Statement Under PCT Article 19

Deleted claim 1,2,3;

Revise

claim

14,15, changed the sequence number and the sequence number that has added accompanying drawing of accompanying drawing.

These are revised specification sheets and accompanying drawing without any influence.

Claims (according to the modification of the 19th of treaty)

1,2,3 (deletions)

4. the electric current by cutting off x axle on the horizontal plane and use the axial electric current of z on the vertical surface, make fragment vertically shift to collection device, wherein on sepharose, flow at electric current on the horizontal plane, and it is the fragment vertical shifting is collected, and a kind of by making it move rather than attempt to collect to help to carry out the equipment of next process at the z direction of principal axis from described collection device by on the z direction of principal axis, applying electric current.

5. the electric current by cutting off x axle on the horizontal plane and use the axial electric current of z on the vertical surface, make fragment vertically shift to collection device, wherein on polyacrylamide gel, flow at electric current on the horizontal plane, and it is the fragment vertical shifting is collected, and a kind of by making it move rather than attempt to collect to help to carry out the equipment of next process at the z direction of principal axis from described collection device by on the z direction of principal axis, applying electric current.

On electric current by cutting off x axle on the horizontal plane and the horizontal plane electric current of y axle and use with two vertical vertical surfaces of line on the axial electric current of z, make fragment vertically shift to collection device, wherein on two-dimentional polyacrylamide gel, flow at electric current on the horizontal plane, and it is the fragment vertical shifting is collected, and a kind of by making it move rather than attempt to collect to help to carry out the equipment of next process at the z direction of principal axis from described collection device by on the z direction of principal axis, applying electric current.

7. by applying electric current at the x of horizontal plane axle with the summation direction of the vertical z axle of x axle, make fragment shift to collection device, wherein on sepharose, flow at electric current on the horizontal plane, similar to the x axle, the y of horizontal plane axle and with the summation direction of the vertical z axle of y axle on apply electric current, make fragment shift to described collection device, and by the x axle, after applying electric current on a certain starting point direction of planar that y axle and z axle form, apply electric current by summation direction at horizontal plane and z axle, make fragment shift to collection device, and mobile fragment collects, and a kind of equipment of carrying out next process rather than attempt to collect from described collection device of helping.

8. by applying electric current at the x of horizontal plane axle with the summation direction of the vertical z axle of x axle, make fragment shift to collection device, wherein on polyacrylamide gel, flow at electric current on the horizontal plane, similar to the x axle, the y of horizontal plane axle and with the summation direction of the vertical z axle of y axle on apply electric current, make fragment shift to described collection device, and by the x axle, after applying electric current on a certain starting point direction of planar that y axle and z axle form, apply electric current by summation direction at horizontal plane and z axle, make fragment shift to described collection device, and mobile fragment collects, and a kind of equipment of carrying out next process rather than attempt to collect from described collection device of helping.

9. form one and have x direction of principal axis and the axial plane of y, wherein on two-dimentional polyacrylamide gel, flow at first electric current on the x direction of principal axis, second electric current flows on the y direction of principal axis, the z axle is perpendicular to other two axle settings, after applying electric current on a certain starting point direction of planar, apply electric current by summation direction at horizontal plane and z axle, make fragment shift to described collection device, and mobile fragment collects, and a kind of equipment of carrying out next process rather than attempt to collect from described collection device of helping.

10. by with the y direction of principal axis of electric current from the x axle steer horizontal plane that uses agarose gel electrophoresis, make fragment shift to collection device, and it is fragment is moved collect, and a kind of by making it move rather than attempt to collect to help to carry out the equipment of next process at the y direction of principal axis from described collection device by on the y direction of principal axis, applying electric current.

11. by with the y direction of principal axis of electric current from the x axle steer horizontal plane that uses polyacrylamide gel electrophoresis, make fragment shift to collection device, and it is fragment is moved collect, and a kind of by moving them at the y direction of principal axis rather than attempting to collect to help to carry out the equipment of next process from described collection device by on the y direction of principal axis, applying electric current.

12. make fragment shift to collection device by apply electric current to collection device, wherein said collection device is used for through using two-dimentional polyacrylamide gel electrophoresis to separate the fragment that obtains, and mobile fragment collects, and a kind of equipment of carrying out next process rather than attempt to collect from described collection device of helping.

13. being held, current direction is used to collect the container of segmental buffered soln or electrolytic solution or the electrode of slit, be immersed in length in buffered soln or the electrolytic solution greater than 0.01mm but less than the electrode of 90mm, be immersed in width in buffered soln or the electrolytic solution greater than 0.01mm but less than the electrode of 90mm.

14. (modification) holds container or the slit that is used to collect segmental buffered soln or electrolytic solution, wherein as Figure 10, Figure 11 and shown in Figure 12, conductor partly is coated with.

15. (modification) held buffered soln or electrolytic solution, is used to collect segmental container or slit, wherein as shown in Figure 7 and Figure 8, film partly is coated with.

Be used to the container or the slit that use electrophoresis apparatus to collect segmental buffered soln or electrolytic solution 16. hold, its wall is wider than 0.01mm but less than 40mm.

17. distance (84) between at the bottom of the hole of collection device and the gel bottom and described collection device are very short to the length (85) (68) bottom the gel from the bottom of y axle bottom.

18. for collection device, must have slit, its length (79) is greater than 0.4mm but less than 40mm, its length (79) greater than bore length (80) 1/5 but less than 40 times of bore length, its width greater than hole width 1/10 but less than 10 times of hole width.

19. can with＜-＜+flow direction of electric current is from x axle steer and the axial electrophoresis chamber of the corresponding y of x axle, wherein for carry out electrophoresis initial setting x axle in the agar electrophoresis groove.

20. in electrophoresis chamber can with＜-＜+flow direction of electric current turns to the axial device of z from x direction of principal axis or y direction of principal axis.

21. for electrophoresis chamber, place the part of gel or gel slab and must be made by transparent material, so that light can penetrate, and rest part is black or Adjacent color, so that light can be partially absorbed.

Claims

1. fragment flows along electric current mobile x direction of principal axis on sepharose, form the hole at the x direction of principal axis, described fragment will be shifted to described hole, and use fragment absorption agent is collected the fragment in the described hole, a kind of collection buffered soln or method of electrolyte, and a kind of method of utilizing rest segment in the described hole to carry out another process.

2. fragment flows along electric current mobile x direction of principal axis on polyacrylamide gel, form the hole at the x direction of principal axis, described fragment will be shifted to described hole, and use fragment absorption agent is collected the fragment in the described hole, a kind of collection buffered soln or method of electrolyte, and a kind of equipment that utilizes rest segment in the described hole to help to carry out next process.

3. fragment flows along electric current mobile x direction of principal axis on two-dimentional polyacrylamide gel, and when electric current fragment when the y direction of principal axis flows also flows, form the hole at the y direction of principal axis, described fragment will be shifted to described hole, use fragment absorption agent is collected the fragment in the described hole, a kind of collection buffered soln or method of electrolyte, and a kind of equipment that utilizes rest segment in the described hole to help to carry out next process.

14. hold the container or the slit that are used to collect segmental buffered soln or electrolytic solution, wherein as shown in Figure 7 and Figure 8, conductor partly is coated with.

15. hold buffered soln or electrolytic solution, be used to collect segmental container or slit, wherein as shown in Figure 5, film partly is coated with.