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中国农学通报

中国农学通报 ›› 2013, Vol. 29 ›› Issue (27): 154-158.doi: 10.11924/j.issn.1000-6850.2012-3500

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

3种椰子基因组DNA提取方法的比较

罗意 黄绵佳 范海阔 肖勇   

  • 收稿日期:2012-10-27 修回日期:2012-12-13 出版日期:2013-09-25 发布日期:2013-09-25

The comparison of three methods for extracting coconut genomic DNA

  • Received:2012-10-27 Revised:2012-12-13 Online:2013-09-25 Published:2013-09-25

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摘要： 旨在得到一套快速提取椰子DNA的方法。采用CTAB小样法、SDS-CTAB法以及PVP法提取椰子组织的DNA，并通过后续的PCR扩增和酶切等实验比较了3种方法所提取的DNA质量的优劣。将CTAB小样法提取的DNA与SDS-CTAB法和PVP法提取的DNA进行了比较，CTAB小样法提取的DNA的OD260/OD280(1.839)较另外2种方法SDS-CTAB法(1.709)和PVP法(1.613)的值更高，CTAB小样法提取的DNA适合于PCR扩增，扩增的条带清晰，且特异性好。此外，CTAB小样法提取的DNA，酶切也较为均匀。CTAB小样法能高效地提取椰子的DNA，并且能一次提取大量的高质量DNA样品，因而能满足高通量的实验要求，同提取DNA能较好地满足下游的分子生物学研究。

关键词: 种植制度, 种植制度

Abstract: The study was carried out for developing a rapid method for DNA of extracting coconut. The quality of DNA extracted with mini-CTAB, SDS-CTAB and PVP method was compared. In this study, the comparison of three sets of coconut DNA, which were isolated separately by mini-CTAB and SDS-CTAB method or PVP method, showed that DNA isolated by mini-CTAB had higher OD260/280 (1.839) than that of SDS-CTAB (1.709) or PVP method (1.613). Meanwhile DNA isolated with the three methods was used for the downstream PCR amplification and restriction analysis. The DNA template from mini-CTAB method produced more clearer and specific bands. The double-enzyme cleavage of DNA from mini-CTAB method showed more even bands. The mini-CTAB method can be used to extract DNA from coconut tissues efficiently and multiple samples with high quality at a time. Since that this method could be used in high throughout experiments which including abundant samples. Furthermore, this method produced DNA which was suitable for downstream molecular biology experiment.