单管双向等位基因专一性扩增方法在近交系小鼠遗传检测中的应用

目的将新近建立的单管双向等位基因专一性扩增(single-tube bi-directional allele specific amplification,SB-ASA)方法用于分析近交系小鼠基因组中的单核苷酸多态性(SNP)。方法以5个近交系小鼠为研究对象,采用SB-ASA方法对其16个SNP位点进行检测,并通过双盲实验和测序验证该方法的可靠性;且考察了该方法中PCR反应各成分及扩增条件对结果的影响。结果16个SNP位点,SB-ASA都成功地对5个品系小鼠进行了分型,与测序结果完全一致;双盲实验结果显示通过3个SNP位点即可鉴别5个品系。结论SB-ASA方法可用于近交系小鼠SNP的遗传检测,可望作为一种新的分子生物学遗传检测方法推广应用。

Application of Single-Tube Bi-Directional Allele Specific Amplification in SNP Genetic Monitoring of Inbred Mice

Objective To analyze single nucleotide polymorphism(SNP) of inbred mouse genes by a lately established method-single-tube bi-directional allele specific amplification(SB-ASA).Method 16 SNP were detected by the SB-ASA method in 5 inbred mouse strains.We also designed a double-blind experiment and sequencing to validate the reliability of this method,and assessed its influence on the results of PCR system and amplification conditions.Results SB-ASA was successfully used to type 5 inbred mouse strains for 16 SNP loci,and the results were completely consistent with those obtained by sequencing.Five inbred mouse strains were successfully distinguished with a double-blind experiment by just 3 SNP loci.Conclusion SB-ASA can be used to SNP genetic monitoring of inbred mice,and it can be popularized and applied as a new molecular biological method of genetic monitoring.