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| 3种椰子基因组DNA提取方法的比较 | |
| 引用本文： | 罗意,黄绵佳,范海阔,肖勇.3种椰子基因组DNA提取方法的比较[J].中国农学通报,2013,29(27):154-158. |
| 作者姓名： | 罗意 黄绵佳 范海阔 肖勇 |
| 作者单位： | 1. 中国热带农业科学院椰子研究所，海南文昌571339; 海南大学园艺园林学院，海口570228 2. 中国热带农业科学院椰子研究所，海南文昌571339; 海南省热带油料作物重点实验室，海南文昌571339 3. 海南大学园艺园林学院,海口,570228 |
| 基金项目： | 中央级公益性科研院所基本科研业务费专项“基于高通量测序的椰子转录组分析”(1630032012028)。 |
| 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： | 椰子 DNA提取 CTAB法 SDS法 PVP法 |
| 收稿时间： | 2012/10/27 0:00:00 |
| 修稿时间： | 2012/12/13 0:00:00 |
The comparison of three methods for extracting coconut genomic DNA |
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| Luo Yi , Fan Haikuo , Huang Mianjia , Xiao Yong.The comparison of three methods for extracting coconut genomic DNA[J].Chinese Agricultural Science Bulletin,2013,29(27):154-158. | |
| Authors: | Luo Yi Fan Haikuo Huang Mianjia Xiao Yong |
| 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. |
| Keywords: | coconut DNA extraction CTAB method SDS method PVP method |
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