Characterization of the synthetic compatible solute homoectoine as a potent PCR enhancer.

نویسندگان

  • Michael Schnoor
  • Peter Voss
  • Paul Cullen
  • Thomas Böking
  • Hans-Joachim Galla
  • Erwin A Galinski
  • Stefan Lorkowski
چکیده

Different substances such as dimethyl sulfoxide, tetramethylene sulfoxide, 2-pyrollidone, and the naturally occurring compatible solute betaine enhance PCR amplification of GC-rich DNA templates with high melting temperatures. In particular, cyclic compatible solutes outperform traditional PCR enhancers. We therefore investigated the effects that cyclic naturally occurring ectoine-type compatible solutes and their synthetic derivatives have on melting temperature of double-stranded DNA (dsDNA) and on PCR amplification of different templates. L-ectoine, betaine, and derivatives of L-ectoine decreased, whereas beta-hydroxyectoine increased, the melting temperature of dsDNA. The ability to decrease the melting temperature was greatest for homoectoine, a new synthetic derivative of l-ectoine. Furthermore, compatible solutes, especially homoectoine, enhanced PCR amplification of GC-rich DNA (72.6% GC content; effective range: 0.1-0.5M).

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عنوان ژورنال:
  • Biochemical and biophysical research communications

دوره 322 3  شماره 

صفحات  -

تاریخ انتشار 2004