[关键词]
[摘要]
分别采用试剂盒和CTAB法提取小麦干种子基因组DNA,DNA样品经紫外光谱吸收、琼脂糖凝胶电泳、PCR扩增检测。结果表明,试剂盒提取DNA操作过程快速简便、成本较低,DNA产率和纯度优于CTAB法。通过多重PCR技术分别以小麦LMW-GS亚基Glu-B3位点和黑麦碱secalin-p基因的特异引物进行扩增,均能够扩增出清晰的目标条带,提取的基因组DNA能够满足分子检测的实验要求。
[Key word]
[Abstract]
Two rapid DNA isolation methods from dry seeds of wheat was established by DNA extraction kit and CTAB in this study. The DNA samples were detected in terms of the UV spectrophotometer analysis, agarose gel electrophoresis, PCR amplification respectively. The results indicats that the method with DNA Extraction Kits was simple, rapid and low costs, the extraction rate and purity of DNA was better than that of the CTAB. Using the DNA samples as template, the Glu-B3 locus of wheat LMW-GS and ω-secalin gene of triticale could be amplified with distinct and specific target bands by multiplex PCR. The genomic DNA extracted from dry seeds of wheat can meet the needs of molecular tests.
[中图分类号]
S512.1;Q781
[基金项目]
甘肃省农业科学院农业科技创新专项“甘肃小麦品种资源1BL/1RS易位系的分子标记检测研究”(2012GAAS06-4)、甘肃省农业生物技术研究与应用开发项目“持久条锈病抗源BJ144抗锈基因发掘及抗锈小麦新品系的培育”(GNSW-2011-07)、甘肃省科技支撑计划项目“小麦5B染色体基因组抗锈基因关联分析及抗锈新品系培育”(1304NKCA142)部分内容