藜麦SSR-PCR反应体系的优化及引物筛选

Optimization of SSR-PCR Reaction System and Primer Selection in Quinoa

藜麦原产南美洲,因其全营养性被誉为"超级谷物"而广受重视,国内对藜麦的研究也逐步开展。本文通过研究SSR-PCR反应体系的五个主要成分对扩增结果的影响,建立了藜麦SSR-PCR反应的优化体系,即总体积10μL:DNA模板75ng,10×PCR buffer(Mg2+plus)1.2μmol/L、dNTP 0.8μmol/L、Taq酶0.2μmol/L,引物1.2μmol/L,余下部分由dd H2O补足。利用优化后的SSR-PCR反应体系对引物进行筛选,从221对引物中成功筛选出条带清晰、特异性好的引物18对。优化后的SSR反应体系和筛选出的引物可为藜麦种质资源的遗传多样性研究、指纹图谱的建立等提供科学依据。

Chenopodium quinoa(quinoa)is native to South America.It is widely regarded as a"super grain"because of its high nutrient value.The domestic research on quinoa is also gradually carried out.In this paper,by studying the influence of five main components of SSR-PCR reaction system on the amplification results,an optimization system of SSR-PCR reaction of quinoa was established.The total volume of SSR-PCR reaction was 10 uL:DNA template 75 ng,10×PCR buffer(Mg2+plus)1.2μmol/L,dNTP 0.8μmol/L,Taq enzyme 0.2μmol/L,primer 1.2μmol/L,and the rest was supplemented by dd H2O.The optimized SSR-PCR reaction system was used to screen the primers,and 18 pairs of primers with clear bands and good specificity were successfully screened from 221 pairs of primers.The optimized SSR reaction system and selected primers can provide scientific basis for the study of genetic diversity and the establishment of fingerprints of quinoa germplasm resources.