A Preliminary Study On Microsatellite Genotyping By Using Hair Shaft

.Genotyping of microsatellite using hair shaft was studied using with reference to the genotyping using skin tissues in a population of 35 individual arctic foxes(Alopex lagopus). DNA was isolated from hair shaft and skin tissues by using routine phenol-chloroform method with a little optimization.Primer pairs were designed for twelve microsatellite loci and five were qualified to apply to the hair shaft genotyping after optimization of PCR conditions. Among the five loci,only AJ411682 is monomorphic.Compared with genotyping results of skin DNA,we and evaluated microsatellite genotype using hair shaft DNA through such indicators as rate of positive amplification,rate of correct genotyping,allelic frequency,genotypic frequency,observed heterozygosity,excepted heterozygosity,matching probability of two random individuals,and discrimination power.Results indicated that the rate of positive amplification of hair shaft at X94309, AJ269545,AY223968 and AJ132981 were 100.00%,80.00%,82.35%,60.00%and 60.00% respectively,and the rate of correct genotyping were 91.43%,74.29%,73.53%,48.00%and 60.00%respectively.There was no significant difference in the observed heterozygosity,excepted heterozygosity,matching probability and discrimination power between skin tissue and hair shaft at 4 polymorphic loci.Accumulated discriminate power was 0.975055097 for skin tissue,and 0.96909861 for hair shaft.The results suggested that(1) hair shaft could be valid for microsatellite genotyping if amplified fragment was not longer than 100 bp;(2) Locus was an impact factor for amplification success;(3) templet quantity recovered from hair shaft was an key impacting factor for microsatellite genotyping;(4) all microsatellites should be fully validated prior to the application to hair shaft genotyping.