Method for high-volume sequencing of nucleic acids: random and d

Details Method for high-volume sequencing of nucleic acids: random and d Method for high-volume sequencing of nucleic acids: random and d

1993-10-12

1995-04-18

Zitomer, Stephanie W.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 911, 935 77, C12Q 168, C12P 1934

Patent

active

054077990

ABSTRACT:
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.

REFERENCES:
patent: 5043272 (1991-08-01), Hartley
Binns et al., J. Virol. Meth. 12:265-269 (1985).
Feinberg et al., Anal. Biochem. 132:6-13 (1983).
Roberts, Science 242:1244-1246 (1988).
Sanger et al., J. Mol. Biol. 143:161-178 (1988).

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