白草种子基因组DNA五种提取方法的比较

Comparison of 5 DNA Extraction Methods from Pennisetum flaccidum Seed

用5种方法 (传统CTAB法、改良CTAB法、高盐低p H法、改良SDS法、试剂盒法)对白草种子基因组DNA进行提取,分别用紫外分光光度计法、琼脂糖凝胶电泳法、PCR扩增检测提取的DNA的质量。结果表明,传统CTAB法提取的DNA浓度低,含有较多杂质,抑制PCR反应;改良SDS法和改良CTAB法提取的DNA纯度低,不符合检测要求;高盐低pH法和试剂盒法提取的DNA纯度高、完整性好,但试剂盒法成本高、DNA得率低。所以高盐低pH法为5种方法中最经济、效果最好的白草种子DNA提取方法。

The objective of this study was to explore a suitable method for genomic DNA extraction from Pennisetum flaccidum seed by comparing five DNA extraction methods （ traditional CTAB, improved CTAB, improved SDS, high salt and low pH, kit method）. The quality of extracted DNA was examined with ultraviolet spectrophotome- try, agarose gel electrophoresis and PCR amplification, respectively. The results showed that the DNA extracted by traditional CTAB contained many PCR inhibitors; the purity of DNA extracted by improved CTAB and SDS did not meet the demand of quality standard of DNA in molecular experiment. High salt and low pH and kit method could extract high-quality and good integrity DNA, but kit method cost too much and the yield of DNA was low. Therefore, high salt and low pH was the most suitable and low cost method of these 5 methods.