Direct sequencing of RAPD fragments using 3'-extended oligonucleotide primers and dye terminator cycle-sequencing.

Abstract:

:Random amplified polymorphic DNA (RAPD) markers are used widely to develop high resolution genetic maps and for genome fingerprinting. Typically, single oligomers of approximately 10 nucleotides are used to PCR amplify characteristic RAPD marker fragments. We describe an efficient method for the direct end-sequencing of gel-purified RAPD fragments using one primer from a set of four 3'-terminal extended (A, T, C or G) oligonucleotides, identical to the RAPD primer but for the single nucleotide extension. Strand-specific DNA sequence could be independently read from each of the RAPD fragments without recourse to strand separation or fragment cloning. Informative RAPD fragments could be readily converted into mapped STS or SCAR loci using this technology. The 3'-extended primers may also be used to amplify independent genomic RAPD markers.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Mitchelson KR,Drenth J,Duong H,Chaparro JX

doi

10.1093/nar/27.19.e28

keywords:

subject

Has Abstract

pub_date

1999-10-01 00:00:00

pages

e28

issue

19

eissn

0305-1048

issn

1362-4962

pii

gnc028

journal_volume

27

pub_type

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