Abstract
All the mRNAs within a cell and their relative levels are indicative of gene expression within that cell, which is essential for its structure and function in its differentiated state. Therefore, methods for the identification of the specific mRNAs and the quantitation of their levels are invaluable tools for understanding gene expression. Due to high endogenous RNase activity within virtually all living cells, the isolation of good quality RNA with minimal degradation is not a trivial task. This protocol outlines a tried and tested methodology for isolating high quality RNA from embryonic kidneys for various applications including microarray analysis and quantitative reverse transcription PCR (qRT-PCR).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism and function. Cell 116: 281–297
Bessonov S, Anokhina M, Will CL, Urlaub H, Lührmann R (2008) Isolation of an active step I spliceosome and composition of its RNP core. Nature 452:846–850
Brunskill EW, Aronow BJ, Georgas K et al (2008) Atlas of gene expression in the developing kidney at microanatomical resolution. Dev Cell 15:781–791
Wang Z, Gerstein M, Snyder M (2009) RNA-Seq: a revolutionary tool for transcriptomics. Nat Rev Genet 10:57–63
D’Alessio G, Riordan JF (1997) Preface. In: D’Alessio G, Riordan JF (eds) Ribonucleases, structures and functions. Academic, San Diego, pp 15–19
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Thowfeequ, S., Michos, O. (2012). Isolation of High Quality RNA from Embryonic Kidney and Cells. In: Michos, O. (eds) Kidney Development. Methods in Molecular Biology™, vol 886. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-851-1_18
Download citation
DOI: https://doi.org/10.1007/978-1-61779-851-1_18
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-850-4
Online ISBN: 978-1-61779-851-1
eBook Packages: Springer Protocols