"Run-off" synthesis and application of defined single-stranded DNA hybridization probes.

Stürzl M, Roth WK (1990)

Publication Status: Published

Publication Type: Journal article

Publication year: 1990

Journal

Analytical Biochemistry Elsevier Masson

Book Volume: 185

Pages Range: 164-9

Journal Issue: 1

DOI: 10.1016/0003-2697(90)90274-d

Abstract

A simple and efficient method for synthesizing radioactively labeled single-stranded DNA hybridization probes with Thermus aquaticus (Taq) DNA polymerase is described. This is done in a "run-off" polymerization with repeated cycles of denaturation, annealing, and extension. It leads to high yields of a single-stranded DNA of defined length (up to 5000 nt), which is labeled to a high specific activity (1.3 x 10(8) cpm/micrograms DNA). These hybridization probes are equally sensitive as nick-translated DNA probes, but strand specific. This was tested by slot blot hybridization with in vitro-transcribed target RNAs and by Northern blotting. The use of single-stranded DNA hybridization probes combines the benefits of DNA stability and single-strand RNA probes.

Authors with CRIS profile

Michael Stürzl Professur für Molekulare und Experimentelle Chirurgie

Involved external institutions

Max-Planck-Institut für Biochemie / Max Planck Institute of Biochemistry DE Germany (DE)

How to cite

APA:

Stürzl, M., & Roth, W.K. (1990). "Run-off" synthesis and application of defined single-stranded DNA hybridization probes. Analytical Biochemistry, 185(1), 164-9. https://doi.org/10.1016/0003-2697(90)90274-d

MLA:

Stürzl, Michael, and Willy Kurt Roth. ""Run-off" synthesis and application of defined single-stranded DNA hybridization probes." Analytical Biochemistry 185.1 (1990): 164-9.

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