title: Single-primer PCR correction: a strategy for false-positive exclusion.
abstract: Polymerase chain reaction (PCR) technology plays an important role in molecular biology research, but false-positive and nonspecific PCR amplification have plagued many researchers. Currently, research on the optimization of the PCR system focuses on double-primer-based PCR products. This research has shown that PCR amplification based on single-primer binding to the DNA template is an important contributing factor to obtaining false-positive results, fragment impurity, and nonspecific fragment amplification, when the PCR conditions are highly restricted during PCR-based target gene cloning, detection of transgenic plants, simple-sequence repeat marker-assisted selection, and mRNA differential display. Here, we compared single- and double-primer amplification and proposed "single-primer PCR correction"; improvements in PCR that eliminate interference caused by single-primer-based nonspecific PCR amplification were demonstrated and the precision and success rates of experiments were increased. Although for some kinds of experiments, the improvement effect of single-primer PCR correction was variable, the precision and success rate could be elevated at 12-50% in our experiment by this way.
authors.affiliation: Biotechnology Center of Jilin Agricultural University, Changchun, PR China.
authors.forename: J
authors.initials: J
authors.lastname: Ma
authors.forename: P W
authors.initials: PW
authors.lastname: Wang
authors.forename: D
authors.initials: D
authors.lastname: Yao
authors.forename: Y P
authors.initials: YP
authors.lastname: Wang
authors.forename: W
authors.initials: W
authors.lastname: Yan
authors.forename: S C
authors.initials: SC
authors.lastname: Guan
mesh_terms: D015139:Blotting, Southern; D017931:DNA Primers / Q000235:genetics*; D012275:Oryza / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D011401:Promoter Regions, Genetic / Q000235:genetics
chemical_list: D017931:DNA Primers