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Isolation of microsatellite markers from Pinus densiflora Sieb. et Zucc. using a dual PCR technique
Authors:A WATANABE  M G IWAIZUMI  M UBUKATA  T KONDO  C LIAN  T HOGETSU
Institution:1. Forest Tree Breeding Center, Ibaraki, 319-1301, Japan,;2. Graduate School of Agricultural and Life Sciences, the University of Tokyo, Tokyo, 113-8657, Japan
Abstract:We developed seven microsatellite loci from Pinus densiflora using a dual polymerase chain reaction (PCR) technique. Of 186 clones from a library based on suppression PCR, 127 contained microsatellite sequences. Of these, 43 candidates were determined sequences of both flanking regions, and 16 regions from this group were chosen as development markers. Seven of these primer pairs successfully amplified polymorphic single loci among 83 resistant trees against pine wood nematode. The observed heterozygosity of the seven microsatellite markers ranged from 0.247 to 0.843. Mendelian inheritance was confirmed using megagametophytes.
Keywords:dual PCR technique  microsatellite  Pinus densiflora
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