The enhancement of PCR amplification of a random sequence DNA library by DMSO and betaine: application to in vitro combinatorial selection of aptamers.
A PCR method for uniform amplification of a random sequence DNA library is described. A combination of 1 M betaine and 5% DMSO improves the PCR amplification by increasing the ratio of full-length products to shortened products, which are a consequence of nonuniform amplification due to stable secondary structures in the templates. This method is expected to be beneficial for obtaining high-affinity aptamers with stable secondary structures.
Kang
Jonghoon
J
Department of Human Biological Chemistry and Genetics, Sealy Center for Structural Biology, The University of Texas Medical Branch, Galveston, TX 77555-1157, USA.
Lee
Myung Soog
MS
Gorenstein
David G
DG
D001622:Betaine / Q000737:chemistry / Q000494:pharmacology*; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D004121:Dimethyl Sulfoxide / Q000737:chemistry / Q000494:pharmacology*; D015723:Gene Library; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012997:Solvents / Q000737:chemistry / Q000494:pharmacology; D013696:Temperature
D012997:Solvents; D001622:Betaine; D004247:DNA; D004121:Dimethyl Sulfoxide