一种提取梨组织高质量基因组DNA的新方法

A New Method for Extraction of High Quality Genomic DNA from Pear

高质量DNA的提取是开展分子生物学研究的重要环节。针对梨组织中含有大量多糖、多酚等影响DNA提取质量的次生代谢产物的特点,本研究以梨叶片、果皮和果肉为材料,开发了一种新的基因组DNA提取方法——N-月桂酰肌氨酸钠法,并分别与常用的试剂盒及CTAB法进行比较分析。对3种方法提取DNA的总量、浓度及OD值等进行检测分析,结果表明:相比试剂盒及CTAB法,N-月桂酰肌氨酸钠法不需水浴加热,步骤少,操作简单,提取的DNA纯度、浓度均有较大提高,并且适用于大片段DNA的提取。因此,N-月桂酰肌氨酸钠法是一种可靠的高效获得梨基因组高质量DNA的新方法。

Extraction of high-quality DNA is an important step in molecular biology research. Abundant of secondary metabolites, such as polysaccharides, polyphenols etc. in the pear tissues can highly affect the extraction of high quality genomic DNA. According to this character, a new genomic DNA extraction method has been developed in this study, named N-Lauroylsarcosine sodium salt method, which was com- pared with the DNA extraction kit and CTAB method by analyzing the total quantity, concentration and OD value of DNA. The results showed that the N-Lauroylsarcosine sodium salt method is more suitable for extraction of large quantity of pear genomic DNA with higher purity and concentration. It is also time- saving with no need of water bath heating, fewer steps, and easier operation. Thus, the N-Lauroylsar- cosine sodium salt DNA extraction method developed in this study is a reliable and high efficient method for extracting high quality genomic DNA of pear.