Apparatus and method for separating polynucleotides and detecting specific polynucleotide sequences

Claim 3

The system of Claim 1, further including sensor means for sensing the presence of radiative energy in said gel matrix" thereby to detect the presence of hybridized polynucleotides therein.

Claim 4

The system of Claim 2, wherein said sensor means comprises means for sensing said radiative energy and converting said radiative energy to spatially defined electronic signals, and means for displaying said signals.

Claim 5

The system of Claim 1, wherein said separation means comprises: a) chamber means adapted to contain an electrophoretic buffer solution and adapted to receive said frame means; said chamber means including a platform for supporting said frame means and electrode means for applying a potential across said frame means, b) variable power supply means coupled to said electrode means, c) means for sensing the temperature of said gel matrix,'and d) power control means responsive to said sensed temperature to vary the voltage applied to said electrode means to optimize the rate of electrophoresis.

Claim 6

The system of Claim 4 wherein said platform for supporting said frame means contains a temperature control platen connected to said variable power supply means; wherein the power supplied to said platen is regulated by said power control means in response to said sensed temperature.

Claim 7

The system of Claim 4,, further including means for filling and draining said chamber means with electrophoretic buffer, wash buffer, acid solution, and/or dye solution from reservoirs.

Claim 8

The system of Claim 4, further including means movably disposed over said frame means for detecting the presence of a dye front and signalling said presence to said control means to cause the power to be removed from said electrodes.

Claim 9

The system of Claim 1, wherein said probe dispensing means comprises a plurality of dispensing heads each containing a probe reservoir; said probe dispensing assembly being movably mounted on said hybridization chamber so as to enable said assembly to move and deposit probe along the length of said frame means.

Claim 10

The system of Claim 1, wherein said hybridization means further includes means for filling and draining of said hybridization chamber with wash buffer, base solution, and/or dye solution from reservoirs.

Claim 11

The system of Claim 1, further including a drying means disposed in said predetermined path for selectively drying said gel matrix under control of said control means.

Claim 12

A rapid system for the separation and identification of polynucleotides; said system comprising electrophoresis means for separating a mixture of polynucleotides according to charge and size in a gel matrix comprising about 0.2 to about 1.5 mm agarose; hybridization means for the in situ hybridization of said separated polynucleotides in said gel matrix with a complementary probe sequence, wherein either said polynucleotide mixture or said complementary probe sequence is a material labelled with a radiative energy-emitting marker; wash means for removing unhybridized labelled material after hybridization; and detector means for detecting the hybridized labelled material, said detector means comprising a scintillator plate adapted to be disposed over said gel matrix after hybridization, means for electrically scanning said plate to spatially determine points of light emanation and to create electrical signals indicative thereof, means for electronically storing said signals, and means for displaying said signals.

Claim 13

A rapid system for the separation and identification of polynucleotides; said system comprising electrophoresis means for separating a mixture of polynucleotides according to charge and size in a gel matrix comprising about 0.2 to about 1.5 mm agarose; hybridization means for the in situ hybridization of said separated polynucleotides in said gel matrix with a complementary probe sequence, wherein either said polynucleotide mixture or said complementary probe sequence is a material labelled with a radiative energy-emitting marker; wash means for removing unhybridized labelled material after hybridization; and detector means for detecting the hybridized labelled material, said detector means comprising a scintillator plate adapted to be disposed over said gel matrix after hybridization, means for electrically scanning said plate to spatially determine points of light emanation and to create electrical signals indicative thereof, means for electronically storing said signals, and means for displaying said signals.

Claim 14

The system of Claim 11, further including microwave drying means for drying said gel matrix.

Claim 17

The apparatus of Claim 13, wherein said platform for supporting said backing means contains a temperature control platen connected to said variable power means; wherein the power supplied to said platen is regulated by said control means in response to said sensed temperature.

Claim 18

The apparatus of Claim 13, further including means for filling and draining said chamber means with electrophoretic buffer, wash buffer, acid solution, and/or dye solution from reservoirs.

Claim 19

The apparatus of Claim 13, further including a dye-front detector means movably disposed over said frame means for detecting the presence of a dye front and signalling said presence to said control means to cause the power to be removed from said electrodes.

Claim 22

The apparatus of Claim 17, wherein said probe dispensing means comprises a plurality of dispensing heads each containing a probe reservoir; said probe dispensing assembly being movably mounted on said hybridization chamber so as to enable said assembly to move and deposit probe along the length of said gel matrix.

Claim 23

The apparatus of Claim 17, further including means for filling and draining said hybridization chamber with wash buffer, base solution, and/or dye solution from individual reservoirs.

Claim 26

A process for the detection of a specific target polynucleotide fragment in a mixture of polynucleotides, said process comprising: a) electrophoretically separating said mixture of polynucleotides on a gel matrix which comprises about 0.2 to about 1.5 mm agarose gel, thereby producing within said gel a series of polynucleotide bands wherein each band represents polynucleotide sequences separated by size and charge, b) adding to said polynucleotide bands which have been previously denatured a polynucleotide probe comprising a polynucleotide sequence the target polynucleotide whose presence or absence is to be determined, wherein either the target polynucleotide or the probe polynucleotide is labelled with a radiative energy-emitting material, c) allowing the .target polynucleotide to rest in contact with the probe polynucleotide until hybridization has occurred, d) removing non-hybridized labelled polynucleotides, and e) analyzing said gel matrix for the presence or absence of hybridized labelled polynucleotides.

Claim 27

A process for the detection of a specific target polynucleotide fragment in a mixture of polynucleotides, said process comprising: a) electrophoretically separating said mixture of polynucleotides on a gel matrix which comprises about 0.2 to about 1.5 mm agarose gel, thereby producing within said gel a series of polynucleotide bands wherein each band represents polynucleotide sequences separated by size and charge, b) adding to said polynucleotide bands which have been previously denatured a polynucleotide probe comprising a polynucleotide sequence the target polynucleotide whose presence or absence is to be determined, wherein either the target polynucleotide or the probe polynucleotide is labelled with a radiative energy-emitting material, c) allowing the .target polynucleotide to rest in contact with the probe polynucleotide until hybridization has occurred, d) removing non-hybridized labelled polynucleotides, and e) analyzing said gel matrix for the presence or absence of hybridized labelled polynucleotides.

Claim 28

The process of Claim 21, wherein said agarose gel comprises about 0.5 to about 2.0% (w/v) agarose.