3 replies to this topic

[Mimoza]

Most people are using the methyltranferase M.SssI to creative fully methylated controls. However, from what I have read, M.SssI methylates only CpG sites. Thus, the rest of Cs of the genome which are not followed by a G remain unmethylated (or have random methylation status depending on the sample). So, this is not a fully methylated control.
Do you know any way to have a truly completely methylated control, independently from the species?
Thanks a lot,
Mimoza

[lmg]

You buy one from Qiagen (human). The prize is worth the effort.

best,
lmg

Edited by lmg, 12 March 2013 - 04:10 AM.

[Mimoza]

Thanks for the answer.
I'm not working with human and I need to have a DNA from my species (2 different fish). Any other idea?

[phage434]

You can PCR with 5-me dCTP instead of dCTP in the reaction. You'll have to mix with dATP, dGTP, and dTTP yourself.