Hi All,
I am performing EMS mutagenesis now and this is my first try. I am wondering how many genes would be hit in my case? I am not familiar about the “math” calculation on EMS treatment… Could anyone help me out?
Basically I am screening aound 30,000 haploid genomes and the EMS concentration I used was 50mM.
Any comments are welcome. Thank you in advance.
Emily
hey emily,
you are going to hit a lot of genes. if your screening assay is efficient you will be approaching saturation for EMS-induced mutations. But you will still miss some things that you could get with ENU.
you should look at phil anderson’s 1995 overview of mutagenesis. a modern day classic. and for the wgs view of things (including spectra for EMS vs ENU vs UV-TMP), see sarin et al. and flibotte et al. good luck with your screen!
eric
Genetics. 2010 June; 185(2): 417–430.
doi: 10.1534/genetics.110.116319
PMCID: PMC2881126
Analysis of Multiple Ethyl Methanesulfonate-Mutagenized Caenorhabditis elegans Strains by Whole-Genome Sequencing
Sumeet Sarin,* Vincent Bertrand,* Henry Bigelow,† Alexander Boyanov, Maria Doitsidou,* Richard J. Poole,* Surinder Narula,* and Oliver Hobert*,1
Genetics: Published Articles Ahead of Print, published on May 3, 2010 as 10.1534/genetics.110.116616
Whole-Genome profiling of mutagenesis in Caenorhabditis elegans.
Stephane Flibotte,* ,§ Mark L. Edgley,** Iasha Chaudhry,** Jon Taylor,** Sarah E. Neil,** Aleksandra Rogula,** Rick Zapf,** Martin Hirst§, Yaron Butterfield§, Steven J. Jones§, Marco M. Marra§, Robert J. Barstead§§ and Donald G. Moerman*,**
Methods in Cell Biology
Volume 48, 1995, Pages 31–58
Chapter 2 Mutagenesis
Philip Anderson
Department of Genetics University of Wisconsin Madison, Wisconsin 53706