3种椰子基因组DNA提取方法的比较

The Comparison of Three Methods for Extracting Genomic DNA of Coconut

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.