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

Aspartate-Assay

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1. Make standards using sodium phosphate at the following uM concentrations: 0, 2, 5, 7, 10, 20, 40, 60, and 80. Use the screw top glass tubes.2. Dry

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实验概要        In certain fruits, such as pineapples and mangoes, the flesh contains protein-digesting enzymes (proteases). These may play a part in

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 This procedure is for cells in 96 well plates, if larger plates are used then adjust volumes accordingly.1 Make a solution of 5mg/ml MTT dissolved in

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PROTOCOL:•Deparaffinize and rehydrate slides:3 x 3´ Xylene3 x 2´ 100% ethanol1 x 2´ 95%, 80%, 70% ethanol (each)1 x 5´ 1x PBS•Microwave antigen retrie

Protease-assay

In certain fruits, such as pineapples and mangoes, the flesh contains protein-digesting enzymes (proteases). These may play a part in helping to softe

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This enzyme is famous for being involved in the development of the GMO tomatoes (more information from the link at the foot of this page). The cells o

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DescriptionVarious phenotypic characteristics are requiredfor a cancer cell to successfully complete the metastaticcascade. Among these, acquisition o

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Bradford-Assay

The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie Brilliant blue

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DescriptionThis is a fast and easy assay to test the angiogenic/anti-angiogenic properties of molecules. As compared to other angiogenesis assays, suc

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DescriptionCancer cells do not show anchorage and contact inhibition of growth. To assess the anchorage and contact independent growth of cells, noble

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Materials to be prepared beforehand:1) FBS free medium2) 10% FBS medium3) Cell migration filter insert ( Transwell®, 12mm Diameter, 12 μm Pore Size.)P

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-Use sterile technique and sterile solutions throughout this method.-1. Grow a starter culture at 30 C with shaking (250 rpm) until it reaches saturat

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