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Gene. 1985;40(2-3):317-23.

Rapid and reliable dideoxy sequencing of double-stranded DNA.

Abstract

We report a simple and reliable protocol for nucleotide sequencing using the Sanger dideoxy technique on linearized double-stranded DNA molecules with specific oligonucleotide primers. The method is demonstrated for restriction fragments cloned into the plasmid vectors pSP64 and pSP65 using two vector-specific primers, the M13 reverse primer and a new SP6 primer, flanking the multiple cloning site. Template DNA may be prepared by a rapid alkaline lysis procedure. Mild linearization conditions with the appropriate restriction endonuclease avoid the appearance of artifact bands.

PMID:
3007296
DOI:
10.1016/0378-1119(85)90055-1
[Indexed for MEDLINE]

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