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Cited by 752

The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.

Direct Cloning and Sequence Analysis of Enzymatically Amplified Genomic Sequences

Scharf, Stephen J., Horn, Glenn T., Erlich, Henry A.·1986·Science

Abstract

A method is described for directly cloning enzymatically amplified segments of genomic DNA into an M13 vector for sequence analysis. A 110-base pair fragment of the human β-globin gene and a 242-base pair fragment of the human leukocyte antigen DQα locus were amplified by the polymerase chain reaction method, a procedure based on repeated cycles of denaturation, primer annealing, and extension by DNA polymerase I. Oligonucleotide primers with restriction endonuclease sites added to their 5′ ends were used to facilitate the cloning of the amplified DNA. The analysis of cloned products allowed the quantitative evaluation of the amplification method's specificity and fidelity. Given the low frequency of sequence errors observed, this approach promises to be a rapid method for obtaining reliable genomic sequences from nanogram amounts of DNA.

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Metadata

Authors
Scharf, S. J., Horn, G. T., & Erlich, H. A.
Venue
Science
Published
1986-09-05
Volume
233
Issue
4768
Pages
1076-1078
ISSN
  • 0036-8075 (ISSN-L)
  • 1095-9203
OpenAlex
W2030903599
Cited by
752
References
1
FWCI
22.66
Type
journal article
Language
en
Open access
closed

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