✦ LIBER ✦

Preparation of single-stranded deoxyribonucleic acid probes using an immobilized template

✍ Scribed by C. Hansen; F. Libert; G. Vassart; D. Christophe

Book ID: 102987554
Publisher: Elsevier Science
Year: 1987
Tongue: English
Weight: 918 KB
Volume: 162
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(87)90017-0

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✦ Synopsis

A new method for the easy preparation of specific single-stranded DNA fragments is presented. Recombinant Ml3 DNA containing the strand complementary to the sequence of interest is made partially double-stranded by elongating a conventional Ml3 sequencing primer. Following linearization by enzymatic digestion downstream from the insert (relative to priming site), this DNA is coupled to diazotized paper through its single-stranded (vector) portion. Subsequent denaturation of the double-stranded region generates an immobilized template strand. Successive runs of primed syntheses of the (desired) complementary strand can be realized using the same template. The copies are easily isolated by release upon denaturation. DNA probes prepared by this method have proven to be valuable tools for gene analysis. Q 1987 Academic Press, Inc.

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