Direct PCR sequencing with boronated nucleotides.

Abstract:

:A method is described to simultaneously amplify and sequence DNA using a new class of nucleotides containing boron. During the polymerase chain reaction, boron-modified nucleotides, i.e. 2'-deoxynucleoside 5'-alpha-[P-borano]-triphosphates, are incorporated into the product DNA. The boranophosphate linkages are resistant to nucleases and thus the positions of the boranophosphates can be revealed by exonuclease digestion, thereby generating a set of fragments that defines the DNA sequence. The boranophosphate method offers an alternative to current PCR sequencing methods. Single-sided primer extension with dideoxynucleotide chain terminators is avoided, with the consequence that the sequencing fragments are derived directly from the original PCR products. Boranophosphate sequencing is demonstrated with the Pharmacia and the Applied Biosystems 373A automatic sequencers, producing data that is comparable with cycle sequencing.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Porter KW,Briley JD,Shaw BR

doi

10.1093/nar/25.8.1611

subject

Has Abstract

pub_date

1997-04-15 00:00:00

pages

1611-7

issue

8

eissn

0305-1048

issn

1362-4962

pii

gka247

journal_volume

25

pub_type

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