title: Characterization of the synthetic compatible solute homoectoine as a potent PCR enhancer.
abstract: 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).
authors.affiliation: Institute of Arteriosclerosis Research, University of Münster, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Schnoor
authors.forename: Peter
authors.initials: P
authors.lastname: Voss
authors.forename: Paul
authors.initials: P
authors.lastname: Cullen
authors.forename: Thomas
authors.initials: T
authors.lastname: Böking
authors.forename: Hans-Joachim
authors.initials: HJ
authors.lastname: Galla
authors.forename: Erwin A
authors.initials: EA
authors.lastname: Galinski
authors.forename: Stefan
authors.initials: S
authors.lastname: Lorkowski
mesh_terms: 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
chemical_list: D000599:Amino Acids, Diamino; D017931:DNA Primers; C121557:hydroxyectoine; D004247:DNA