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模板DNA的制备是制约分子育种效率的瓶颈步骤之一。本研究介绍一种高通量快速的玉米基因组DNA提取方法,即蔗糖提取液法。该方法切取20~30 mg玉米种子胚乳,通过批量化研磨、加热和瞬时离心3个步骤,即可获得稳健PCR扩增的模板DNA,并且在光温培养箱条件下切取部分胚乳的剩余种子与对照完整种子发芽率无显著差异。通过蔗糖提取液法提取192粒F2种子胚乳DNA,鉴定WxWx、Wxwx和wxwx 3种基因型与表型鉴定结果相一致。蔗糖提取液法提取胚乳DNA简单、快速、成本低廉,可有效用于高通量分子标记辅助选择。
The preparation of template DNA is one of the bottleneck steps to restrict the efficiency of molecular breeding. This study introduced a high-throughput method for rapid extraction of maize genomic DNA, namely sucrose extract method. The method comprises the following steps of: cutting the endosperm of 20-30 mg corn seeds and obtaining stable template DNA amplified by batch milling, heating and instantaneous centrifugation, and cutting the remaining seeds of partial endosperm in a light-temperature incubator with There was no significant difference in the germination rate of the control intact seeds. Two hundred and seventy-two F2 endosperm DNA samples were extracted by sucrose extract method, and the three genotypes of WxWx, Wxwx and wxwx were identified consistent with phenotypic identification. Sucrose extract method extraction of endosperm DNA is simple, rapid, low cost, which can be effectively used for high-throughput molecular marker-assisted selection.