Method for high-volume sequencing of nucleic acids: random and directed priming with libraries of oligonucleotides
Random and directed priming methods for determining nucleotide sequences by enzymatic sequencing techniques, using libraries of primers of lengths 8, 9 or 10 bases, are disclosed. These methods permit direct sequencing of nucleic acids as large as 45,000 base pairs or larger without the necessity for subcloning. Individual primers are used repeatedly to prime sequence reactions in many different nucleic acid molecules. Libraries containing as few as 10,000 octamers, 14,200 nonamers, or 44,000 decamers would have the capacity to determine the sequence of almost any cosmid DNA. Random priming with a fixed set of primers from a smaller library can also be used to initiate the sequencing of individual nucleic acid molecules, with the sequence being completed by directed priming with primers from the library. In contrast to random cloning techniques, a combined random and directed priming strategy is far more efficient. 2 figs.
- Issue Date:
- OSTI Identifier:
- 46303
- Assignee:
- Associated Universities, Inc., Washington, DC (United States) PTO; SCA: 550200; PA: EDB-95:079160; SN: 95001384534
- Patent Number(s):
- US 5,407,799/A/
- Application Number:
- PAN: 8-135,317
- Contract Number:
- AC02-76CH00016
- Resource Relation:
- Other Information: PBD: 18 Apr 1995
- Research Org:
- Associated Universities Inc
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 55 BIOLOGY AND MEDICINE, BASIC STUDIES; DNA SEQUENCING; MOLECULAR STRUCTURE; DNA-CLONING; MOLECULAR BIOLOGY; ENZYMES