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Isolation of Total RNA from Tissues or Cell Lines

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RNA Isolation and Characterization Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 86))

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Abstract

The purity and integrity of Isolated RNA IS a critical determinant of its effectiveness in such molecular biological procedures as Northern blot, poly A+ RNA separation, cDNA synthesis, and in vitro transcription and translation The successful isolation of total RNA from tissues or cell lines by any 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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References

  1. Chirgwin J. M., Przybyla A. E., MacDonald R. J., and Rutter W. J. (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Blochem J 18, 5294.

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  2. Chomczynski P. and Sacchi N (1987) Methods of RNA isolation by acid guam-dinium thiocyanate-phenol-chloroform extraction Anal Biochem 162, 156–159.

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© 1998 Humana Press Inc.

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Mukhopadhyay, T., Roth, J.A. (1998). Isolation of Total RNA from Tissues or Cell Lines. In: Rapley, R., Manning, D.L. (eds) RNA Isolation and Characterization Protocols. Methods in Molecular Biology™, vol 86. Humana Press. https://doi.org/10.1385/0-89603-494-1:55

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  • DOI: https://doi.org/10.1385/0-89603-494-1:55

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-494-5

  • Online ISBN: 978-1-59259-570-9

  • eBook Packages: Springer Protocols

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