RNA酶保护试验((RNaseProtectionAssay,RPA)简介
RNA酶保护试验((RNase Protection Assay,RPA)是通过液相杂交的方式,用反义RNA探针与样品杂交,以检测RNA表达的技术。1。原理:双链RNA(杂交的)能够抵抗RNA酶的降解。2。应用:检测RNA表达3。与Northern杂交和RT-PCR比较,RPA有以下几个优点:1. 检测灵敏度比Northern杂交高。由于Northern杂交步骤中转膜和洗膜都将造成样品和探针的损失,使灵敏度下降,而RPA将所有杂交体系进行电泳,故损失小,提高了灵敏度。2. 由于PCR扩增过程中效率不均一和反应“平台”问题,基于PCR产物量进行分析所得数据的可靠性将下降,而RPA没有扩增过程,因此,分析的数据真实性较高。3. 由于与反义RNA探针杂交的样品RNA仅为该RNA分子的部分片段,因此,部分降解的RNA样品仍可进行分析。4. 步骤较少,耗时短。与Northern杂交相比,省去了转膜和洗膜的过程。5. RNA-RNA杂交体稳......阅读全文
The-ribonuclease-protection-assay-(RPA)
The ribonuclease protection assay (RPA) is a highly sensitive and specific method for the detection of mRNA species. The assay was made possible by th
Roche公司的RNase-Protection-Assay-(RPA)-protocol
Roche公司的RNase Protection Assay (RPA) Using DIG-Labeled RNA Probes下载网址:http://www.roche-applied-science.com/PROD_INF/BIOCHEMI/no1_03/PDF/p22_23.pdf还有一份
RNA酶保护实验(RNase-Protection-Assay,RPA)简介
简介: RNA酶保护试验(RNase Protection Assay,RPA)是通过液相杂交的方式,用反义RNA探针与样品杂交,以检测RNA表达的技术。 1. 原理:双链RNA(杂交的)能够抵抗RNA酶的降解。 2. 应用:检测RNA表达 3. 与Northern杂交和RT-PCR比较
RNA酶保护实验(RNase-Protection-Assay,RPA)简介
简介:RNA酶保护试验(RNase Protection Assay,RPA)是通过液相杂交的方式,用反义RNA探针与样品杂交,以检测RNA表达的技术。1. 原理:双链RNA(杂交的)能够抵抗RNA酶的降解。2. 应用:检测RNA表达3. 与Northern杂交和RT-PCR比较,RPA有以下几个优
RNA酶保护试验((RNase-Protection-Assay,RPA)简介
RNA酶保护试验((RNase Protection Assay,RPA)是通过液相杂交的方式,用反义RNA探针与样品杂交,以检测RNA表达的技术。1。原理:双链RNA(杂交的)能够抵抗RNA酶的降解。2。应用:检测RNA表达3。与Northern杂交和RT-PCR比较,RPA有以下几个优点:1.
RNA酶保护试验((RNase-Protection-Assay,RPA)方法
一、试剂准备1. GACU POOL:取100mM ATP、CTP、GTP各2.78μl、100mM UTP 0.06μl,加DEPC H2O至100μl。2. 杂交缓冲液IPES 0.134g、0.5M EDTA(pH8.0)20μl、5M NaCl 0.8ml、甲酰胺8ml,加DEPC H2O至
RNA酶保护试验((RNase-Protection-Assay,RPA)的优缺点
RNA酶保护试验((RNase Protection Assay,RPA)是通过液相杂交的方式,用反义RNA探针与样品杂交,以检测RNA表达的技术。与Northern杂交和RT-PCR比较,RPA有以下几个优点:1. 检测灵敏度比Northern杂交高。由于Northern杂交步骤中转膜和洗膜都将造
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