Isolation and characterization of 16 microsatellite loci for the giant salamander Andrias davidianus
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MICROSATELLITE LETTERS
Isolation and characterization of 16 microsatellite loci for the giant salamander Andrias davidianus Jie Wang • Jing Zhang • Xulin Li • Jianping Jiang
Received: 3 November 2013 / Accepted: 25 November 2013 / Published online: 5 December 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract Sixteen microsatellite markers were developed for the critically endangered giant salamander Andrias davidianus to examine its population genetic structure and parentage across central China. The number of alleles per locus ranged from 4 to 10 (mean of 7.4) when tested on 36 individuals from two cultivated populations (LY, TB). The average observed and expected heterozygosities in LY (n = 19) and TB (n = 17) were 0.83 ± 0.02 and 0.79 ± 0.01, 0.70 ± 0.03 and 0.67 ± 0.02 respectively. Keywords Andrias davidianus Microsatellite Mating system Population genetic structure
Chinese giant salamander (CGS) Andrias davidianus is the world’s largest amphibian species and it is classified as critically endangered in the Chinese Red Book of Amphibians and Reptiles and on the IUCN Red List. Its current distribution is highly fragmented and some wild populations are already extirpated (Wang et al. 2004). Because of its biological, medical, and nutritional value, it has been cultured widely in many provinces of China for more than 30 years without reports of its mating system. Here we described 16 microsatellite loci for A. davidianus, which
Electronic supplementary material The online version of this article (doi:10.1007/s12686-013-0095-2) contains supplementary material, which is available to authorized users. J. Wang J. Zhang X. Li J. Jiang (&) Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, Sichuan, China e-mail: [email protected] J. Wang e-mail: [email protected]
will enable investigation of the mating system and population genetic structure of this species. A total of 36 samples of larvae were collected in two cultivated farms of CGS that located in Lueyang County (33.343 N, 106.353 E, 600 m a.s.l.) and Taibai County (33.725 N, 107.429 E, 850 m a.s.l.), respectively. Genomic DNA was extracted from muscle tissues using TransGen EasyPure Genomic DNA Extraction Kit. An enriched partial genomic library for the repeat motifs (GATA)n were obtained following essentially the Fast Isolation by AFLP of Sequences Containing Repeats (FIASCO) protocol (Zane et al. 2002). One hundred and ninety clones were sequenced, of which 88 ones had simple repeating motifs and adequate flanking regions and could be used to create primer pairs. A total of 16 loci were successfully amplified in the 36 individuals and the forward primers were labeled with fluorescent dye FAM, HEX, TAM or ROX. The amplification reaction used 10 lL PCR mixture containing approximately 0.7 ng of genomic DNA, 0.4 lM of each primer and 4 lL EasyTaq PCR SuperMix, and performed using the following cycling profiles: initial denaturation at 94 °C for 3 min; followed by 35 cycles at 94 °C for 30 s, at annealing temperatu
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