The mRNA Decay Resource:Protocols and Resources
Protocols
Yeast protocols
From the Atkin lab
Total RNA extraction Support Protocol: Preparation of buffer saturated phenol
Measurement of Saccharomyces cerevisiae mRNA half lives
Kebaara et al., 2006. Determination of mRNA half-lives in Candida albicans using thiolutin as a transcription inhibitor. Genome. 49(8):894-9. [1]
Yeast polyribosome preparation for RNA
Electrophoresis of RNA through gels containing formaldehyde
Northern transfer of RNA
Preparation of oligolabeled probes
Hybridization of RNA blots
Removal of oligolabeled probes from GeneScreen Plus membranes
Yeast polyribosome preparation - Proteins
Gel electrophoresis of proteins in denaturing conditions
Electrotransfer of proteins from polyacrylamide gels to membranes
Immunoblotting
From the Jacobson lab
He et al., 2008. Qualitative and quantitative assessment of the activity of the yeast nonsense-mediated mRNA decay pathway. Methods Enzymol. 449:127-47. [2]
From Fred Sherman
Sherman, 2002. Getting started with yeast. Methods Enzymol. 350:3-41. [3]
From Mike Culbertson
Global measurement of mRNA half lives. Guan et al., 2006. Impact of nonsense-mediated mRNA decay on the global expression profile of budding yeast. PLoS Genet. 2(11):e203. Epub 2006 Oct 18. [4]
General RNA Protocols
Capped RNAs:
Bajak and Hagedorn, 2008. Efficient 5' cap-dependent RNA purification: use in identifying and studying subsets of RNA. Methods Mol Biol. 419:147-60. [5]
Blewett et al., 2011. A quantitative assay for measuring mRNA decapping by splinted ligation reverse transcription polymerase chain reaction: qSL-RT-PCR. RNA 17(3):535-43. Epub 2011 Jan 10. [6]
