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

• Published in: Aquaculture
• Main Authors: Li, Ronghua, Li, Qi, Cornette, Florence, Degremont, Lionel, Lapegue, Sylvie
• Format: Article in Journal/Newspaper
• Language: English
• Published: Elsevier Science Bv 2010
• Subjects: Pacific oyster; Crassostrea gigas; Microsatellite; EST; Multiplex PCR; Parentage assignment; Pacific
• Online Access: https://archimer.ifremer.fr/doc/00013/12419/9999.pdf; https://doi.org/10.1016/j.aquaculture.2010.09.037; https://archimer.ifremer.fr/doc/00013/12419/

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.