Development of four EST-SSR multiplex PCRs in the Pacific oyster (Crassostrea gigas) and their validation in parentage assignment

Type Article
Date 2010-12
Language English
Author(s) Li Ronghua1, 2, Li Qi2, Cornette Florence1, Degremont LionelORCID1, Lapegue Sylvie1
Affiliation(s) 1 : IFREMER, Lab Genet & Pathol, F-17390 La Tremblade, France.
2 : Ocean Univ China, Coll Fisheries, Qingdao 266003, Peoples R China.
Source Aquaculture (0044-8486) (Elsevier Science Bv), 2010-12 , Vol. 310 , N. 1-2 , P. 234-239
DOI 10.1016/j.aquaculture.2010.09.037
WOS© Times Cited 23
Keyword(s) Pacific oyster, Crassostrea gigas, Microsatellite, EST, Multiplex PCR, Parentage assignment
Abstract We report four highly informative multiplex PCRs developed from twelve previously described ESTSSRs in Crassostrea gigas. We evaluated and validated these multiplex PCRs in twelve full-sib families. The average allelic richness and the polymorphism information content (PIC) were 11.1 and 0.811 respectively. The combined power of exclusion was greater than 99.99% using all four multiplex assays. A hundred and forty three tests of segregation ratios revealed 11 significant departures from expected Mendelian ratios. The frequency of null alleles was estimated as 4.9% of all the alleles segregating based on a within-family analysis of Mendelian segregation patterns. Parentage analysis of real offspring demonstrated that 97% of all offspring were unambiguously allocated to a pair of parents based on two multiplex PCRs with only a 4% error rate, and 100% of the offspring were correctly allocated to their parents when three multiplex PCRs were used.
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