4种高梁基因组DNA快速提取方法的比较

摘要

[目的]寻找快速、简单、成本低的高粱基因组DNA提取方法.[方法]首先,分别使用液氮研磨法、缓冲液研磨法、烘干研磨法和直接研磨法这4种方法从高粱叶片中提取基因组DNA,然后,通过凝胶电泳、SSR分析和SRAP分析检测所提DNA的浓度和纯度.[结果]采用4种方法提取高粱基因组DNA时的产量相差不大;采用前2种方法提取的DNA降解少,纯度高,可进行有效的SRAP-PCR和SSR-PCR,但缓冲液研磨法的SRAP-PCR结果稍差;而采用后两种方法提取的DNA降解严重,纯度低,但可进行有效的SSR-PCR.[结论]采用4种方法均可在短时间内提取到足够几十次PCR反应的高粱基因组DNA,液氮研磨法和缓冲液研磨法所提取的DNA应用范围更大,而烘干研磨法和直接研磨法所提取的DNA只能用于对片段较小的目的DNA进行特异性PCR.%[Objective] This study was to find out a quick, simple, and low-cost method for the extraction of sorghum genomic DNA. [Method] Four plant genomic DNA extraction methods based on CTAB, including liquid nitrogen grinding method ( method Ⅰ ), buffer grinding method (method Ⅱ), drying grinding method (method Ⅲ ) and directly grinding method (method Ⅳ), were used to extract the sorghum genomic DNA from leaves; further the quantity and quality of the yielded DNA were detected by gel electrophoresis, SSR-PCR and SRAP-PCR. [Result] These four methods performed no remarkable difference in DNA product. The method Ⅰ and method Ⅱ produced DNA with higher purity and better integrity, which, especially from method Ⅰ, is effective for SRAP-PCR and SSR-PCR. While the DNA extracted via method Ⅲ and method Ⅳ had less integrality and lower purity, and only effective in SSR-PCR. [Conclusion] Enough amount of sorghum genomic DNA to perform tens of PCR could be quickly extracted using all these four methods. The DNA obtained via method Ⅰ and method Ⅱ had a broader application spectrum (SRAP, RAPD, ISSR and SSR) than that via method Ⅲ and method Ⅳ which is only proper for PCR targeting small DNA fragments (SSR).