摘要： 以13个河北核桃优系(罗汉头、南安庄、艺核1号、C12、J16、M3、H12、C15、M10、MB、C13、M9、A7)的叶片为材料,比较了高盐低pH法(2%PVP40,3%PVP40)、SDS法(0.5%,1%,1.5%)和CTAB法(2%,3%)提取基因组DNA的效果,旨在筛选出适合河北核桃DNA提取的最佳方法,为进一步开展河北核桃的分子群体遗传研究奠定基础。通过紫外分光光度计法和琼脂糖凝胶电泳等方法对所提取的DNA样品进行检测,比较所得DNA的纯度和质量。结果表明,几种提取方法均能提取到核桃叶片的基因组DNA,其中含3%PVP40的高盐低pH法所提取的DNA浓度适中,蛋白质、RNA、多糖等去除彻底,无降解且OD260/OD280值为1.86,确定3%PVP40的高盐低pH法为河北核桃叶片DNA提取的最适方法。
关键词:
河北核桃,
叶片,
基因组DNA,
提取,
方法
Abstract: Using the leaves of Juglans hopeiensis Hu(Luohantou,Nananzhuang,Yihe1,C12,J16,M3,H12,C15,M10,MB,C13,M9,A7)as the experimental materials,three extraction methods of genomic DNA including high-salt and low-pH(2%PVP40,3%PVP40),SDS(0.5%,1%,1.5%)and modified CTAB(2%,3%)were compared,in order to select the optimal DNA extraction method and establish base of molecular population genetics of Juglans hopeiensis.The DNA samples were tested by agarose gel electrophoresis and ultraviolet spectrophotometer.The results showed that several extraction methods were able to extract genomic DNA,but there were some differences in purity and quality.3% PVP40 high-salt and low-pH method was the optimal method by compairisng purity and quality of the DNA.
Key words:
JuglanshopeiensisHu,
Leaf,
GenomicDNA,
Extraction,
Method
中图分类号:
雷玲, 王红霞, 张志华, 何富强, 高仪, 赵书岗, 孙红川. 河北核桃叶片DNA提取方法的比较[J]. 华北农学报, 2010, 25(2): 145-148. doi: 10.7668/hbnxb.2010.02.029.
LEI Ling, WANG Hong-xia, ZHANG Zhi-hua, HE Fu-qiang, GAO Yi, ZHAO Shu-gang, SUN Hong-chuan. Comparison of DNA Extraction Methods from Leaves of Juglans hopeiensis Hu[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2010, 25(2): 145-148. doi: 10.7668/hbnxb.2010.02.029.