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。
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 OD2JODzs0 (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.
出处
《中国农学通报》
CSCD
2013年第27期154-158,共5页
Chinese Agricultural Science Bulletin
基金
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)