Direct sequencing of unpurified PCR-amplified DNA by semi-exponential cycle sequencing (SECS).

Molecular biotechnology ·Vol. 8 ·No. 3 ·1997-12-00 ·Pages 269-77

Sarkar G, Bolander ME

Abstract

A simple technique for direct sequencing of PCR-amplified templates without purification of the PCR reaction product is presented. This method does not require an additional synthesis step after template amplification, and can generate sequence information form as little as 0.1 fmol of unpurified template.

MeSH Terms

DNA/analysis,isolation & purification Polymerase Chain Reaction/methods Sequence Analysis, DNA Templates, Genetic

Chemicals

DNA

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Sarkar G

Mayo Clinic, Rochester, MN 55905, USA. [email protected]

Bolander M E

References (11)

11 references, click to expand

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Rapid sequencing of unpurified PCR products by thermal asymmetric PCR cycle sequencing using unlabeled sequencing primers.
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A system for shotgun DNA sequencing.
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Rapid one-step characterization of recombinant vectors by direct analysis of transformed Escherichia coli colonies.
Biotechniques. 1989 Jul-Aug;7(7):689-90 PMID: 2517212
DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA.
Proc Natl Acad Sci U S A. 1988 Dec;85(24):9436-40 PMID: 3200828
DNA sequencing with chain-terminating inhibitors.
Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
Restriction-site PCR: a direct method of unknown sequence retrieval adjacent to a known locus by using universal primers.
PCR Methods Appl. 1993 May;2(4):318-22 PMID: 8391890
A rapid polymerase-chain-reaction-directed sequencing strategy using a thermostable DNA polymerase from Thermus flavus.
Gene. 1992 Apr 1;113(1):17-23 PMID: 1563631
Sequencing products of the polymerase chain reaction directly, without purification.
Biotechniques. 1990 Feb;8(2):142-8 PMID: 2317367
Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction.
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Article Info

Journal

Molecular biotechnology

Abbr.

Mol Biotechnol

ISSN

1073-6085

DOI

10.1007/BF02760780

[email protected]

Published

1997-12-00

Pages

269-77

Language

English

Region

United States

NLM ID

9423533

Subset

External Links

PubMed source DOI: 10.1007/BF02760780 View journal details

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