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Titlebook: RNA Isolation and Characterization Protocols; Ralph Rapley,David L. Manning Book 1998 Springer Science+Business Media New York 1998

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發(fā)表于 2025-3-21 16:15:40 | 只看該作者 |倒序瀏覽 |閱讀模式
書目名稱RNA Isolation and Characterization Protocols
編輯Ralph Rapley,David L. Manning
視頻videohttp://file.papertrans.cn/821/820174/820174.mp4
叢書名稱Methods in Molecular Biology
圖書封面Titlebook: RNA Isolation and Characterization Protocols;  Ralph Rapley,David L. Manning Book 1998 Springer Science+Business Media New York 1998
描述Ribonucleic acids are central to cellular and molecular processes and perform vital functions in both structural and functional roles. RNA molecules form the bridge between the stable genetic information contained within DNA and enzymes and proteins that carry out much of the metabolism within the cell. Many of the sites of protein synthesis, the ribosomes within the cell, are composed of these ribonucleic acids as are the tRNA molecules that deliver the amino acid building blocks to the ribosomes. Of all the RNA species, the nucleic acid intermediate, messenger RNA, is a desirable source of material to biologists, since this reflects much of, what ultimately, is translated into enzymes and proteins. In order to determine the qualitative and quantitative changes in mRNA expression, a vast number of molecular biological techniques have been developed. Key molecular methods that provide the means to initially isolate and analyze RNA molecules are the focus of this volume. In putting together this collection of protocols, we have tried to provide techniques that are most applicable and widely used. In particular, there are a number of iso- tion techniques included that have been devel
出版日期Book 1998
版次1
doihttps://doi.org/10.1385/0896034941
isbn_softcover978-1-4899-4250-0
isbn_ebook978-1-59259-570-9Series ISSN 1064-3745 Series E-ISSN 1940-6029
issn_series 1064-3745
copyrightSpringer Science+Business Media New York 1998
The information of publication is updating

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Nonradioactive Northern Blotting,nt of chemiluminescent substrates (e.g., CSPD, CPD, CPD-Star from Troptx) has improved the sensitivity of the detection to a range that was not achieved with calorimetric detection via chromogenic substrates (e.g., 5-chloro-4-bromo-indolylphosphate) for alkaline phosphatase.
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Isolation of Total RNA from Tissues or Cell Lines, procedure involves four major steps: complete disruption of the cells or tissues; effective denaturation of the nuclear protein complex; inactivation of endogenous ribonuclease (RNase) activity; and purification of the RNA from the contaminating DNA, protein, and carbohydrates.
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Formaldehyde Gel Electrophoresis of Total RNA,ently, the electrophoresis of RNA needs to be performed under denaturing conditions. Heat denaturing the RNA sample prior to electrophoresis is insufficient, as secondary structures will simply reform unless a denaturing system is used.
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1064-3745 olecules form the bridge between the stable genetic information contained within DNA and enzymes and proteins that carry out much of the metabolism within the cell. Many of the sites of protein synthesis, the ribosomes within the cell, are composed of these ribonucleic acids as are the tRNA molecule
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An Improved Rapid Method of Isolating RNA from Cultured Cells,l RNA. Most protocols rely on sodium dodecyl sulfate (SDS) (.) or guamdium thiocyanate (.,.) to simultaneously lyse cells and mactivate endogenous ribonucleases. In those procedures, the RNA 1s separated from cellular DNA and proteins by centrifugation through 5.7 . CsC1, or by acid phenol extractio
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Isolating RNA with the Cationic Surfactant, Catrimox-14,phase, and the cationic head groups bind to the nucleic acid electrostatically. The precipitate can be dissolved in organic solvents, or it can be redissolved in water by the addition of salt. Selected cationic surfactants are capable of lysing cells, by solubilizmg their protem and lipid components
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