Site-Specific PCR for Distinguishing Bubali Cornu from Adulterants
Objective:To develop a polymerase chain reaction(PCR)-based method for the rapid and accurate distinguishing of Bubali Cornu from adulterants.Methods:The specific single nucleotide polymorphisms(SNPs)of Bubalus bubalis Linnaeus were screened by comparison on the cytochrome C oxidase subunit Ⅰ(COⅠ)gene sequences between Bubalus bubalis,Bos taurus,Bos mutus,and Capra hircus,on the basis of which corresponding primers were designed.The PCR system was optimized,and the tolerability and applicability of the method were evaluated.Results:The PCR conditions were as follows:the amplification system of 20 μL,annealing temperature of 60℃,32 cycles,DNA template of 100 ng,0.8 μL forward and reverse primers(10 μmol·L-1 each).The amplification of Bubali Cornu samples was carried out with the designed primer SN1,which yielded a specific band at approximately 358 bp after agarose gel electrophoresis,while the samples from other species did not show any band.Conclusion:The developed PCR method enables rapid and accurate distinguishing of Bubali Cornu from other common animal horn samples.This method serves as a basis for distinguishing Bubali Cornu from adulterants.
万方数据
维普
