Characterization of the synthetic compatible solute homoectoine as a potent PCR enhancer.
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).
Schnoor
Michael
M
Institute of Arteriosclerosis Research, University of Münster, Germany.
Voss
Peter
P
Cullen
Paul
P
Böking
Thomas
T
Galla
Hans-Joachim
HJ
Galinski
Erwin A
EA
Lorkowski
Stefan
S
D000599:Amino Acids, Diamino / Q000494:pharmacology*; D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000187:drug effects; D017931:DNA Primers; D004305:Dose-Response Relationship, Drug; D007700:Kinetics; D009691:Nucleic Acid Denaturation / Q000187:drug effects; D016133:Polymerase Chain Reaction; D013698:Templates, Genetic
D000599:Amino Acids, Diamino; D017931:DNA Primers; C121557:hydroxyectoine; D004247:DNA