骆驼刺叶片总RNA提取方法比较  

Comparison of Total RNA Isolating Methods from Alhagi Sparsifolia Leaves

作　　者：努尔凯麦尔.木拉提 王希东[1] 帕尔哈提.阿布都克日木 

机构地区：[1]新疆农业大学农学院,乌鲁木齐830052 [2]喀什师范学院生物与地理科学系,新疆喀什844008

出　　处：《喀什师范学院学报》2013年第6期44-46,共3页Journal of Kashgar Teachers College

摘　　要：骆驼刺具有抗寒、抗旱、耐盐和抗风沙的特性,是一种非常好的生物性抗逆研究材料.通过比较CTAB法、热硼酸法和Trizol法来提取骆驼刺叶片总RNA的效果,结果表明,Trizol试剂盒法提取的RNA纯度较高,电泳条带清晰完整,OD360/OD280比值为1.91,并将其确定为骆驼刺叶片总RNA提取的方法,反转录合成cDNA第一链,利用设计的NHX1引物,PCR扩增得到一条600～800bp的DNA条带,与报道的NHX1基因的核心序列大小基本相符,验证了提取的RNA可用于RT-PCR,为骆驼刺的抗逆性研究奠定基础.Alhagi sparsifolia is a very good biological resistance materials with the characteristics of cold resistance, drought resistance, salt resistance and wind characteristics. In order to find out a method of isolation of total RNA,isolated the total RNAfromthe Alhagi sparsifolia's leaves used CTABmethod, hot borate method and Trizol kit method, the result proved the Trizol kit method is the best for its purity is higher, the belt is clear and integrated, the rate of OD260 /OD280 is 1.91. Then designed the special primers to synthesis the 1st cDNA, and amplified by PCR, gained a DNA belt about 600 bp to 800 bp, which conforms to the core sequence of the NHX1 gene that had published. It proved the RNA can be applied to RTPCR, and lay a foundation for the molecular biology research works of the resistance aspects on Alhagi sparsifolia.

关 键 词：骆驼刺 CTAB 热酚法 TRIZOL RT-PCR 

分 类 号：Q943[生物学—植物学]