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DEVELOPMENT OF MULTIPLEX PCR OF MICROSATELLITES AND ITS APPLICATION TO KINSHIP ANALYSIS IN CRASSOSTREA GIGAS
Author(s): 
Pages: 1542-1548
Year: Issue:  6
Journal: Oceanologia et Limnologia Sinica

Keyword:  Crassostrea gigasmicrosatellitemultiplex PCRparentage analysis;
Abstract: We tested two sets of high-level 5-multiplex PCRs for efficient genotyping in the Pacific oyster(Crassostrea gigas). Ten loci were selected based on polymorphism, reliability, and allele size range. They were divided into two groups according to allele size and labeled with four different fluorescence dyes, so that loci in the same color were not be overlap in size. In addition, we performed a paternity test for 27 full-sib families and 643 offspring of Group 0527 and 27 full-sib families and 382 offspring of Group 0612 with software CERVUS 3.0. Parentage analysis demonstrated that 100% of all offspring were allocated unambiguously to the unique pair of parents when two sets of 5-multiplex PCRs were used for Groups 0527 and 0612. The assignment success for the two groups was 96% based on MS 1(the first 5-multiplex PCR). However, the assignment success was 97% and 95% based on MS 2(the second 5-multiplex PCR) for Groups 0527 and 0612 respectively. Therefore, the two sets of 5-multiplex PCRs can be used for rapid and efficient parentage assignment in the Pacific oyster.
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