Apparatus and method for the generation, separation, detection, and recognition of biopolymer fragments

• US 5,993,634 A
• Filed: 05/09/1996
• Issued: 11/30/1999
• Est. Priority Date: 05/09/1995
• Status: Expired due to Fees

• Current Assignee: CuraGen Corporation (Celldex Therapeutics Incorporated)
• Original Assignee: CuraGen Corporation (Celldex Therapeutics Incorporated)
• Inventors: Went, Gregory T., Rothberg, Jonathan M., Simpson, John W.
• Primary Examiner(s): McKane, Elizabeth
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US08/644,073
• Time in Patent Office: 1,300 Days
• Field of Search: 204/612, 204/603, 204/461, 204/452, 204/605, 204/604, 204/602, 204/601, 204/600, 204/450, 204/451, 204/453, 204/454, 204/455, 204/616, 702/19, 702/20, 356/344, 356/328
• US Class Current: 204/612
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/00783 Laminate assemblies, i.e. the reactor comprising a stack of plates
• CPC Class Codes: B01J 2219/00828 Silicon wafers or plates
• CPC Class Codes: B01J 2219/00831 Glass
• CPC Class Codes: B01J 2219/00833 Plastic
• CPC Class Codes: B01J 2219/00873 Heat exchange
• CPC Class Codes: B01J 2219/00995 Mathematical modeling
• CPC Class Codes: G01N 27/44708 Cooling
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G01N 27/44782 of a plurality of samples
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

This invention is an integrated instrument for high-capacity electrophoretic analysis of biopolymer samples. It comprises a specialized high-voltage, electrophoretic module in which the migration lanes are formed between a bottom plate and a plurality of etched grooves in a top plate, the module permitting concurrent separation of 80 or more separate samples. In thermal contact with the bottom plate is a thermal control module incorporating a plurality of Peltier heat transfer devices for the control of temperature and gradients in the electrophoretic medium. Fragments are detected by a transmission imaging spectrograph which simultaneously spatially focuses and spectrally resolves the detection region of all the migration lanes. The spectrograph comprises a transmission dispersion element and a CCD array to detect signals. Signal analysis comprises the steps of noise filtering, comparison in a configuration space with signal prototypes, and selection of the best prototype. Optionally post-processing is done by a Monte-Carlo simulated annealing algorithm to improve results. Optionally, an array of micro-reactors can be integrated into the instrument for the generation of sequencing reaction fragments directly from crude DNA samples.

Claims

1. An integrated apparatus for concurrent preparation and analysis of a plurality of biopolymer fragment samples, each sample comprising a plurality of fragments obtained from one or more biopolymers, the apparatus comprising:

(a) means for substantially concurrent electrophoretic separation of each of a plurality of biopolymer fragment samples loaded into an electrophoretic separation medium;

(b) means for substantially simultaneously stimulating light emissions from fragments in a plurality of biopolymer fragment samples;

(c) means for substantially simultaneous resolution of said light emissions into spatial and spectral components and generation of output signals representative thereof; and

(d) means for loading the plurality of biopolymer fragment samples into the electrophoretic separation medium comprising

(i) a plurality of wells in the electrophoretic separation medium from which the biopolymer fragment samples migrate for separation, each well containing a buffer medium;

(ii) a solid phase loading comb having a plurality of teeth, each tooth being spaced and sized to fit into one of the plurality of wells, the teeth having means for the adhesion of the biopolymer fragment samples, the adhesion being such that the fragment samples are released upon insertion into the buffer medium in the wells,

said solid phase loading comb being guided into the sample wells by notches formed on a plate, the notches being sized to match the teeth of the comb.

2. An integrated apparatus for concurrent preparation and analysis of a plurality of biopolymer fragment samples, each sample comprising a plurality of fragments obtained from one or more biopolymers, the apparatus comprising:

(a) means for substantially concurrent electrophoretic separation of each of a plurality of biopolymer fragment samples loaded into an electrophoretic separation medium;

(b) means for substantially simultaneously stimulating light emissions from fragments in a plurality of biopolymer fragment samples;

(c) means for substantially simultaneous resolution of said light emissions into spatial and spectral components and generation of output signals representative thereof; and

(d) means for loading the plurality of biopolymer fragment samples into the electrophoretic separation medium comprising;

(i) a plurality of wells in the electrophoretic separation medium from which the biopolymer fragment samples migrate for separation, each well containing a buffer medium; and

(ii) a solid phase loading system comprising a plurality of magnetic beads, such beads being placed into the plurality of wells, the beads having means for adhesion of the biopolymer fragment samples, the adhesion being such that the fragment samples are released upon insertion into the buffer medium in the wells.

3. An integrated apparatus for concurrent preparation and analysis of a plurality of biopolymer fragment samples, each sample comprising a plurality of fragments obtained from one or more biopolymers, the apparatus comprising:

(a) means for substantially concurrent electrophoretic separation of each of a plurality of biopolymer fragment samples loaded into an electrophoretic separation medium;

(b) means for substantially simultaneously stimulating light emissions from fragments in a plurality of biopolymer fragment samples;

(c) means for substantially simultaneous resolution of said light emissions into spatial and spectral components and generation of output signals representative thereof; and

(d) means for loading the plurality of biopolymer fragment samples into the electrophoretic separation medium,

wherein the means for loading further comprises a sample focusing electrode and a means for controlling voltage applied to said electrode for the purpose of concentrating loaded samples.

4. An integrated apparatus for concurrent preparation and analysis of a plurality of biopolymer fragment samples, each sample comprising a plurality of fragments obtained from one or more biopolymers, the apparatus comprising:

(a) means for substantially concurrent electrophoretic separation of each of a plurality of biopolymer fragment samples loaded into an electrophoretic separation medium;

(b) means for substantially simultaneously stimulating light emissions from fragments in a plurality of biopolymer fragment samples;

(c) means for substantially simultaneous resolution of said light emissions into spatial and spectral components and generation of output signals representative thereof; and

(d) means for preparing from an input sample of a plurality of biopolymers a plurality of biopolymer fragment samples for subsequent analysis.

5. An integrated apparatus for concurrent preparation and analysis of a plurality of biopolymer fragment samples, each sample comprising a plurality of fragments obtained from one or more biopolymers, the apparatus comprising:

(a) an electrophoresis module for substantially concurrent electrophoretic separation of each of a plurality of biopolymer fragment samples loaded into an electrophoretic separation medium comprising

(i) a substantially flat bottom plate, and

(ii) a substantially flat top plate having a plurality of grooves on one side, the top plate being positioned in contact with the bottom plate so that the grooves form with the bottom plate a plurality of separated channels, the channels holding the electrophoretic medium to form electrophoretic migration lanes;

(b) means for substantially simultaneously stimulating light emissions from fragments in a plurality of biopolymer fragment samples; and

(c) means for substantially simultaneous resolution of said light emissions into spatial and spectral components and generation of output signals representative thereof;

wherein the grooves in the top plate are spaced to converge toward one end of the plate.

6. An integrated apparatus for concurrent preparation and analysis of a plurality of biopolymer fragment samples, each sample comprising a plurality of fragments obtained from one or more biopolymers, the apparatus comprising:

(a) an electrophoresis module for substantially concurrent electrophoretic separation of each of a plurality of biopolymer fragment samples loaded into an electrophoretic separation medium comprising

(i) a substantially flat bottom plate, and

(ii) a substantially flat top plate having a plurality of grooves on one side, the top plate being positioned in contact with the bottom plate so that the grooves form with the bottom plate a plurality of separated channels, the channels holding the electrophoretic medium to form electrophoretic migration lanes;

(b) means for substantially simultaneously stimulating light emissions from fragments in a plurality of biopolymer fragment samples;

(c) means for substantially simultaneous resolution of said light emissions into spatial and spectral components and generation of output signals representative thereof;

(d) cross-lane grooves between selected adjacent lanes forming cross-lane connecting channels, the channels holding separation medium so that biopolymer fragments can migrate between the selected adjacent lanes through the channels; and

(e) electrodes formed in the walls of the selected lanes adjacent to the cross-lane connecting channels for causing fragment migration through the connecting channels upon being energized with voltage.

7. An integrated apparatus for concurrent preparation and analysis of a plurality of biopolymer fragment samples, each sample comprising a plurality of fragments obtained from one or more biopolymers, the apparatus comprising:

(a) means for substantially concurrent electrophoretic separation of each of a plurality of biopolymer fragment samples loaded into an electrophoretic separation medium;

(b) means for substantially simultaneously stimulating light emissions from fragments in a plurality of biopolymer fragment samples; and

(c) a transmission imaging spectrograph for substantially simultaneous resolution of said light emissions into spatial and spectral components and generation of output signals representative thereof comprising

(i) an optic assembly positioned to receive a substantial fraction of the plurality of light emissions for simultaneous spatial imaging along a first axis and spectral dispersion along a second axis, and

(ii) a detector array for simultaneous spatial and spectral detection of the plurality of light emissions imaged along the first axis and dispersed along the second axis by the optic assembly, the detector producing output signals representative of the detected light;

(iii) a collection lens positioned to initially receive and collimate the plurality of light emissions;

(iv) a transmission dispersion element for spectrally dispersing the collimated signals along the first axis; and

(v) a focusing lens for spatially focusing the signals along the second axis onto the detector array.

8. The apparatus of claim 7 further comprising a spectral filter element positioned between the collection lens and transmission dispersion element for filtering from the light emissions extraneous light.

9. The apparatus of claim 7 in which the transmission dispersion element is a transmission diffraction grating, or a transmission grating-prism.

10. An integrated apparatus for concurrent preparation and analysis of a plurality of biopolymer fragment samples, each sample comprising a plurality of fragments obtained from one or more biopolymers, the apparatus comprising:

(a) means for substantially concurrent electrophoretic separation of each of a plurality of biopolymer fragment samples loaded into an electrophoretic separation medium;

(b) means for substantially simultaneously stimulating light emissions from fragments in a plurality of biopolymer fragment samples;

(c) means for substantially simultaneous resolution of said light emissions into spatial and spectral components and generation of output signals representative thereof; and

(d) means for loading the plurality of biopolymer fragment samples into the electrophoretic separation medium comprising

(i) an array of micro-reactors in which biopolymer fragment samples are generated from biopolymer samples, each micro-reactor having a minute inlet for loading a biopolymer sample;

(ii) a plurality of capillary inlet passages, each micro-reactor having an inlet passage, through which reagents needed for fragment generation are loaded;

(iii) a plurality of capillary outlet passages, each micro-reactor having an outlet passage, through which biopolymer fragment samples are ejected into the electrophoresis module for separation; and

(iv) a plurality of capillary controllers, one controller in each capillary inlet and outlet passage, for controlling fluid flow in the capillaries.

11. The apparatus of claim 10 wherein the capillary controller for controlling fluid flow in a capillary comprises:

(a) an electrical micro-heating element in thermal contact with the capillary; and

(b) electrical leads to the heating elements for energizing the heating elements, whereby current in the leads to a micro-heating element causes fluid evaporation in the contacted capillary and formation of a vapor bubble which blocks fluid flow in the capillary.

12. The apparatus of claim 11 wherein the micro-heating elements comprise:

(a) a layer of a resistive material deposited adjacent to the contacted capillary; and

(b) a layer of protective material deposited over the layer of resistive material for separating the resistive layer from the capillary contents.

13. An integrated apparatus for concurrent preparation of analysis of a plurality of biopolymer fragment samples, each sample comprising a plurality of fragments obtained from one or more biopolymers, wherein the biopolymer samples are DNA samples, the biopolymer fragment samples are DNA sequencing reaction fragments labeled with dyes, each dye having distinctive spectral properties, the apparatus comprising:

(a) means for substantially concurrent electrophoretic separation of each of a plurality of biopolymer fragment samples loaded into an electrophoretic separation medium;

(b) means for substantially simultaneously stimulating light emissions from fragments in a plurality of biopolymer fragment samples;

(c) means for substantially simultaneous resolution of said light emissions into spatial and spectral components and generation of output signals representative thereof;

(d) memory means for storing the output signals of the means for resolution and detection and for storing a set of prototype signals;

(e) means for cumulating the stored output signals into spectral samples, each such spectral sample representative of the distinctive spectral characteristics of the dye labels of one biopolymer fragment sample;

(f) means for comparing the time behavior of the spectral samples with the time behaviors of the set of prototype signals and for selecting prototypes from the set that most closely match the spectral samples; and

(g) means for outputting identities of the selected prototypes as the DNA sequences of the DNA samples.

14. The apparatus of claim 13 wherein the set of prototype signals comprises the output from the analysis of well known DNA sequences.

15. The apparatus of claim 13 wherein the selected prototypes represent pairs of triples of sequential DNA bases.

16. The apparatus of claim 13 wherein the means for selecting comprises comparing a distance metric between the time behaviors of the spectral samples and the time behaviors of the prototypes and selecting as representative that prototype with the closest distance.

17. The apparatus of claim 16 wherein the distance metric is output as an indication of the probable accuracy of the DNA sequences.

18. The apparatus of claim 16 wherein the distance metric is the sum of the squares of the differences in signal values between the spectral samples and the prototype signals.

19. The apparatus of claim 13 further comprising means for outputting identities of multiple samples and relationships between said sample identities.

20. The apparatus of claim 13 further comprising:

(a) means for trimming from the output DNA sequences of the DNA samples stored in memory known DNA sequences in the DNA sample;

(b) means for proofreading in a Monte Carlo manner the trimmed DNA sequences stored in memory, the means for proofreading comprising means for repetitively making at a ransom point in the trimmed output a random sequence alteration and evaluating sequence improvement until no further substantial sequence improvement occurs; and

(c) means for storing and outputting the improved sequences.

21. The apparatus of claim 20 wherein sequence improvement is evaluated by evaluating a probabilistic Boltzman condition on the difference in two distance metrics, one distance metric being between the original DNA sequence and the spectral samples, the other distance metric being between alternative DNA sequences and the spectral samples.

22. An integrated apparatus for concurrent preparation and analysis of a plurality of biopolymer fragment samples, each sample comprising a plurality of fragments obtained from one or more biopolymers, wherein the biopolymer samples are DNA samples, the biopolymer fragment samples are DNA sequencing reaction fragments labeled with dyes, each dye having distinctive spectral properties, the apparatus comprising:

(a) means for substantially concurrent electrophoretic separation of each of a plurality of biopolymer fragment samples loaded into an electrophoretic separation medium;

(b) means for substantially simultaneously stimulating light emissions from fragments in a plurality of biopolymer fragment samples;

(c) means for substantially simultaneous resolution of said light emissions into spatial and spectral components and generation of output signals representative thereof;

(d) memory means for storing the output signals of the means for resolution and detection and for storing a set of prototype signals;

(e) means for cumulating the stored output signals into a time series of spectral samples, each such spectral sample representative of the distinctive spectral characteristics of the dye labels of one biopolymer fragment sample;

(f) means for comparing at a plurality of successive observation times the time behavior of the time series of spectral samples with the time behaviors of the set of prototype signals and for selecting a prototype from the set that most closely matches the spectral samples; and

(g) means for outputting the identity of the prototype that is the closest match.

23. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separation occurring along migration lanes in said electrophoresis module;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial bins representative of said migration lanes and spectral bins representative of said light emissions from said biopolymer fragments, said spatial and spectral bins being adjustable; and

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals;

wherein said biopolymer is DNA and said biopolymer fragment samples are generated in DNA fingerprinting reactions.

24. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separation occurring along migration lanes in said electrophoresis module;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial bins representative of said migration lanes and spectral binds representative of said light emissions from said biopolymer fragments, said spatial and spectral bins being adjustable; and

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals;

wherein said biopolymer is DNA and said biopolymer fragment samples are generated in DNA sequencing reactions.

25. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separation occurring along migration lanes in said electrophoresis module;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial bins representative of said migration lanes and spectral binds representative of said light emissions from said biopolymer fragments, said spatial and spectral bins being adjustable; and

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals;

wherein said means for stimulating light emissions illuminates and fragment samples at multiple wavelengths.

26. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separation occurring along migration lanes in said electrophoresis module;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial bins representative of said migration lanes and spectral binds representative of said light emissions from said biopolymer fragments, said spatial and spectral bins being adjustable; and

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals;

wherein said means for stimulating light emissions comprises a laser.

27. The apparatus of claim 26 wherein said laser is a solid state laser.

28. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separation occurring along migration lanes in said electrophoresis module;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial binds representative of said migration lanes and spectral bins representative of said light emissions from said biopolymer fragments, said spatial and spectral bins being adjustable; and

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals;

wherein said biopolymer fragments are labeled with one or more fluorescent labels, and wherein said means for stimulating light emissions stimulates said light emissions by causing fluorescence of said fluorescent labels.

29. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separation occurring along migration lanes in said electrophoresis module;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial bins representative of said migration lanes and spectral bins representative of said light emissions from said biopolymer fragments, said spatial and spectral bins being adjustable; and

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals;

wherein said biopolymer fragments have distinguishable dye labels from which said light emissions are stimulated, and wherein said spectral bins are adjusted according to the light emission maxima of said dye labels.

30. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separation occurring along migration lanes in said electrophoresis module, said biopolymer fragments having distinguishable dye labels from which said light emissions are stimulated;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial bins representative of said migration lanes and spectral bins representative of said light emissions from said biopolymer fragments, said spatial and spectral bins being adjustable; and

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals, said processor comprising;

(i) a memory for storing said grouped signals and said plurality of event prototypes;

(ii) means for outputting identities of selected event prototypes; and

(iii) means for outputting identities of multiple fragment samples and relationships between said fragment sample identities;

wherein said processor further provides for (1) filtering and converting said stored grouped signals into a plurality of further signals, said further signals representative of the dye label concentrations, for (2) comparing the time behavior of said further signals with the time behaviors of said plurality of event prototypes, and for (3) selecting one or more event prototypes closest to said further signals.

31. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separation occurring along migration lanes in said electrophoresis module, said biopolymer fragments having distinguishable dye labels from which said light emissions are stimulated;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial bins representative of said migration lanes and spectral bins representative of said light emissions from said biopolymer fragments, said spatial and spectral bins being adjustable; and

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals, said processor comprising;

(i) a memory for storing said grouped signals and said plurality of event prototypes; and

(ii) means for outputting identities of selected event prototypes;

wherein said processor further provides for (1) filtering and converting said stored grouped signals into a plurality of further signals, said further signals representative of the dye label concentrations, for (2) comparing the time behavior of said further signals with the time behaviors of said plurality of event prototypes, and for (3) selecting one or more event prototypes closest to said further signals;

wherein said biopolymer is DNA, and wherein said plurality of event prototypes is generated from analyses of known DNA.

32. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separation occurring along migration lanes is said electrophoresis module, said biopolymer fragments having distinguishable dye labels from which said light emissions are stimulated;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial bins representative of said migration lanes and spectral bins representative of said light emissions from the biopolymer fragments, said spatial and spectral bins being adjustable; and

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals, said processor comprising;

(i) a memory for storing said grouped signals and said plurality of event prototypes; and

(ii) means for outputting identities of selected event prototypes;

wherein said processor further provides for (1) filtering and converting said stored grouped signals into a plurality of further signals, said further signals representative of the dye label concentrations, for )2) comparing the time behavior of said further signals with the time behaviors of said plurality of event prototypes, for (3) providing for a distance between said time behaviors of said further signals and said time behaviors of said event prototypes, for (4) selecting one or more event prototypes closest to said further signals, and for (5) selecting those one or more event prototypes with substantially smallest distances; and

wherein said smallest distances are output as an indication of the probable accuracy of the DNA sequences.

33. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separating occurring along migration lanes in said electrophoresis module;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial bins representative of said migration lanes and spectral bins representative of said light emissions from said biopolymer fragments, said spatial and spectral bins being adjustable;

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals; and

(e) a focusing electrode to which a voltage is applied in order to concentrate said biopolymer fragment samples at said focusing electrode prior to the initiation of electrophoresis.

34. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separation occurring along migration lanes in said electrophoresis module, said electrophoresis module comprising;

(i) a first substantially flat plate; and

(ii) a second substantially flat plate having a plurality of non-intersecting grooves on one side, said first plate being positioned in contact with said second plate so that said grooves form with said first plate a plurality of separated channels, each channel suitable for holding an electrophoretic medium and for forming one of said migration lanes;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial bins representative of said migration lanes and spectral bins representative of said light emissions from said biopolymer fragments, said spatial and spectral bins being adjustable; and

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals;

wherein said second plate is borofloat glass and the lanes are formed by etching with hydrofluoric acid.

35. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separation occurring along migration lanes in said electrophoresis module, said electrophoresis module comprising;

(i) a first substantially flat plate; and

(ii) a second substantially flat plate having a plurality of non-intersecting grooves on one side, said first plate being positioned in contact with said second plate so that said grooves form with said first plate a plurality of separated channels, each channel suitable for holding an electrophoretic medium and for forming one of said migration lanes;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial bins representative of said migration lanes and spectral bins representative of said light emissions from said biopolymer fragmetns, said spatial and spectral bins being adjustable; and

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals;

wherein said second plate is cast in a mold.

36. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separation occurring along migration lanes in said electrophoresis module, said electrophoresis module comprising;

(i) a first substantially flat plate; and

(ii) a second substantially flat plate having a plurality of non-intersecting grooves on one side, said first plate being positioned in contact with said second plate so that said grooves form with said first plate a plurality of separated channels, each channel suitable for holding an electrophoretic medium and for forming one of said migration lanes;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial bins representative of said migration lanes and spectral bins representative of said light emissions from said biopolymer fragments, said spatial and spectral bins being adjustable; and

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals;

wherein said second pate is disposable.

37. An integrated apparatus for concurrent separation and analysis of a plurality of biopolymer fragment samples, each said sample comprising a plurality of biopolymer fragments, the apparatus comprising:

(a) an electrophoresis module for the substantially concurrent electrophoretic separation of said biopolymer fragments within each of said plurality of biopolymer fragment samples, said separation occurring along migration lanes in said electrophoresis module, said electrophoresis module comprising;

(i) a first substantially flat plate; and

(ii) a second substantially flat plate having a plurality of non-intersecting grooves on one side, said first plate being positioned in contact with said second plate so that said grooves form with said first plate a plurality of separated channels, each channel suitable for holding an electrophoretic medium and for forming one of said migration lanes;

(b) means for substantially simultaneously stimulating light emissions from said biopolymer fragments;

(c) a transmission imaging spectrograph for producing grouped signals from substantially simultaneous resolution of said light emissions from said biopolymer fragments into spatial binds representative of said migration lanes and spectral bins representative of said light emissions from said biopolymer fragments, said spatial and spectral bins being adjustable; and

(d) a processor for comparing said grouped signals with a plurality of event prototypes and for selecting the one or more event prototypes closest to said grouped signals;

wherein said grooves are formed in a polymeric substrate.