title: An alternative suite of universal primers for genotyping in multiplex PCR.
abstract: The universal primer three-primer approach can dramatically reduce the cost when genotyping the microsatellites. One former research reported four universal primers that can be used in singleplex and multiplex genotyping. In this study, we proposed an alternative suite of universal primers with four dyes for genotyping 8-12 loci in one single run. This multiplex method was tested on Tetranychus truncatus. Published microsatellite loci of T. kanzawai, Frankliniella occidentalis and Nilaparvata lugens were modified as needed and also tested. The robustness of the method was confirmed by comparing with singleplex using multiple fluorophores and genotyping two populations of T. truncatus. This method showed lower signal strength than the singleplex three-primer system, but it was still sufficient to determine the fragment length. The cost of such a project can be reduced dramatically when many loci of different species are involved. In this way, laboratories performing population genetic analyses or studying several different species may benefit from the use of this cost-effective protocol.
authors.affiliation: Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Ge
authors.affiliation: Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.
authors.forename: Yu-Nan
authors.initials: YN
authors.lastname: Cui
authors.affiliation: Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.
authors.forename: Peng-Yu
authors.initials: PY
authors.lastname: Jing
authors.affiliation: Department of Entomology, Nanjing Agricultural University, Nanjing, Jiangsu, China.
authors.forename: Xiao-Yue
authors.initials: XY
authors.lastname: Hong
mesh_terms: D000818:Animals; D017931:DNA Primers; D005260:Female; D056426:Genetic Loci; D060005:Genotyping Techniques / Q000379:methods*; D018895:Microsatellite Repeats; D008925:Mites / Q000145:classification / Q000235:genetics; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers