TY - CHAP
T1 - Determining if an mRNA is a Substrate of Nonsense-Mediated mRNA Decay in Saccharomyces cerevisiae
AU - Johansson, Marcus J O
PY - 2017
Y1 - 2017
N2 - Nonsense-mediated mRNA decay (NMD) is a conserved eukaryotic quality control mechanism which triggers decay of mRNAs harboring premature translation termination codons. In this chapter, I describe methods for monitoring the influence of NMD on mRNA abundance and decay rates in Saccharomyces cerevisiae. The descriptions include detailed methods for growing yeast cells, total RNA isolation, and Northern blotting. Although the chapter focuses on NMD, the methods can be easily adapted to assess the effect of other mRNA decay pathways.
AB - Nonsense-mediated mRNA decay (NMD) is a conserved eukaryotic quality control mechanism which triggers decay of mRNAs harboring premature translation termination codons. In this chapter, I describe methods for monitoring the influence of NMD on mRNA abundance and decay rates in Saccharomyces cerevisiae. The descriptions include detailed methods for growing yeast cells, total RNA isolation, and Northern blotting. Although the chapter focuses on NMD, the methods can be easily adapted to assess the effect of other mRNA decay pathways.
KW - Blotting, Northern
KW - Gene Expression Profiling/methods
KW - Gene Expression Regulation, Fungal
KW - Half-Life
KW - Nonsense Mediated mRNA Decay
KW - RNA, Fungal/genetics
KW - RNA, Messenger/genetics
KW - Saccharomyces cerevisiae/genetics
KW - Saccharomyces cerevisiae Proteins
U2 - 10.1007/978-1-4939-6518-2_13
DO - 10.1007/978-1-4939-6518-2_13
M3 - Book chapter
C2 - 27832540
SN - 1-4939-6516-8
T3 - Methods in molecular biology (Clifton, N.J.)
SP - 169
EP - 177
BT - Eukaryotic Transcriptional and Post-Transcriptional Gene Expression Regulation
ER -