Large scale production of DNA probes
AssigneeMolecular Diagnostics, Inc.
InventorsNanibhushan Dattagupta, Peter M. M. Rae, Donald M. Crothers, Thomas R. Barnett
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Abstract
A process for production of a single strand of a nucleic acid comprising covalently linking to a solid substrate a polynucleotide complementary to the desired strand, hybridizing said polynucleotide with an oligonucleotide, extending the oligonucleotide in direction away from said substrate, denaturing the hybridized polynucleotide and extended oligonucleotide, thereby to free the extended oligonucleotide from the solid substrate, and separating the extended oligonucleotide. The product can be used for making analytical probes.
Claims9
A process for the production of a single strand of a nucleic acid, the processing comprising
- (a) tailing a 3' end of a nucleic acid with a terminal deoxynucleotidyl transferase and a ribonucleoside triphosphate,
- (b) oxidizing the product of step (a) to produce a nucleic acid with a pendant aldehyde group,
- (c) reacting the product the step (b) with a solid support, said solid support containing one or more NH.sub.2 groups,
- (d) reacting the rpduct of step (c) with a primer nucleic acid to form a hybrid,
- (e) extending the primer on the hybrid with DNA polymerase and a nucleoside triphosphate,
- (f) denaturing the product of step (e), and
- (g) separating the product of step (f) into a soluble product and an insoluble product.
Dependencies are inferred from the claim text.