一种快速检测岷江百合总RNA样本中基因组DNA残留的方法  

A Rapid Detection Method of Residual Genomic DNA in Total RNA Samples from Lilium regale

作　　者：杜文凯[1] 袁素霞[2] 胡凤荣[1] DU Wen-kai;YUAN Su-xia;HU Feng-rong(College of Landscape Architecture,Nanjing Forestry Universit,Nanjing 210037;The Institute of Vegetables and Flowers in Chinese Academy of Agricultural Sciences,Beijing 100081)

机构地区：[1]南京林业大学风景园林学院,南京210037 [2]中国农业科学院蔬菜花卉研究所,北京100081

出　　处：《生物技术通报》2018年第10期81-86,共6页Biotechnology Bulletin

基　　金：中国农业科学院科技创新工程(CAAS-ASTIP-IVFCAAS);农业部园艺作物生物学与种质创制重点实验室项目

摘　　要：总RNA样本中残留基因组DNA会严重影响qRT-PCR的准确性。为了检测RNA样本中的DNA残留,依据持家基因TIP41-like的部分内含子序列设计了1对基因组DNA残留检测引物:LDRG-F:5'-CTCTTTTATGACGAGGTTGGTA-3'及LDRG-R:5'-CAGGGAAATCGGTCAGTGTT-3'。利用这对引物对3种不同RNA提取试剂盒提取的总RNA样本以及2种不同第1链c DNA合成试剂盒合成的cDNA样本进行PCR扩增检测,能高效快速地检测岷江百合总RNA以及cDNA样本中有无基因组DNA残留。The residual genomic DNA in total RNA seriously affects the detection accuracy of qRT-PCR.In order to detect residual DNA in the RNA sample,a pair of primers LDRG-F:5'-CTCTTTTATGACGAGGTTGGTA-3'and LDRG-R:5'-CAGGGAAATCGGTCAGTGTT-3'were designed based on partial sequence of introns in the house-keeping gene lTIP41-like.Then this pair of primers were used to detect the total RNA samples extracted by using three different RNA extraction kits and the cDNA samples obtained by using two different first strand cDNA synthesis kits via PCR amplification.The result showed that using the primers helped to rapidly detect whether or not there were the residual genomic DNA in total RNA and cDNA samples from Lilium regal.

关 键 词：岷江百合 基因组DNA残留 TIP41-like 总RNA 检测 

分 类 号：S682.29[农业科学—观赏园艺]