摘要
消减杂交差异显示分析方法(subtractivehybridizationdifferencedisplay,SHDD)是近年来在cD-NA代表性差异分析方法(cDNARepresentationalDifferenceAnalysis,cDNARDA)和mRNA差异显示分析方法(mRNADifferentialDisplay,mRNADD)的基础上发展而来的一种克隆基因的新方法。SHDD方法结合了cDNARDA的特异性特点与mRNADD在双向筛查差异片段和较高的检测灵敏度方面的优势。在具体技术路线设计上,SHDD最大的特点就是在两样本间双向一轮与二轮均衡、温和消减杂交基础上引入了同位素mRNADD差异显示技术,使得该方法在克隆低丰度信息及较弱差异信息、提高差异分析结果的特异性、减少假阳性、减少重复差异结果,提高差异分析的信息量同时又降低工作量等方面明显优于经典的mRNADD和cDNARDA方法。
SHDD （subtractive hybridization difference display） is a newly developed gene-cloning method, which combines a highly specific subtracfive hybridization of cDNA representational difference analysis （cDNA RDA） with a sensitive bi-directional radioactive detection of mRNA differential display （mRNA DD）. The major characteristic of this modified method is adopting a first round of bi-directional subtractive hybridization between two sample cDNAs and an appropriate second round of bi-directional subtractive hybridization between the two resultant first-round difference products, followed by labeling resultant difference products with isotope and separating these difference products on DNA sequencing gel. All these modifications facilitate SHDD method in isolating mRNA species with very low expression levels and/or very small expression differences between two related sample cDNAs, and obtaining more difference products with increased specificity and work etticiency, compared with typical mRNA DD and cDNA RDA.
出处
《国际遗传学杂志》
CAS
2006年第3期183-186,共4页
International Journal of Genetics
基金
国家自然科学基金(No.30400542)
关键词
消减杂交差异显示分析
cDNA代表性差异分析
mRNA差异显示分析
Subtractive hybridization differential display（SHDD）
cDNA representational difference analysis（eDNA RDA）
mRNA differential display（mRNA DD）
