MethylCode Bisulfite Conversion Kit from Invitrogen - Used this kit? (May/14/2009 )
Did anyone use the MethylCode Bisulfite Conversion Kit? Any Comment?
Thank you so much.
I guess it is a new kid on the block. I think the most important parameter measuring the quality of bisulfite conversion kit is DNA recovery.
pcrman on May 22 2009, 05:36 AM said:
I think DNA recovery is only an important parameter for measuring the 'quality' of a bisulfite-conversion kit in case of a small amount of starting material. Okay, I won't use a kit for which I obtain a recovery less than 50 percent. But in my opinion there are more important parameters like for instance:
* complete bisulfite conversion
* low DNA degradation
* reliable results
* short hands-on-time
* DNA input volume
* no residual contaminations
I agree with MoB. Those parameters are very important too. but I think all bisulfite kits are pretty much the same (chemicals), those parameters largely depend on how you implement the protocol, while method for subsequent DNA purification and recovery provided the kits is very different and thus is an important determinant of the kit's quality.
pcrman on May 27 2009, 08:38 AM said:
Bisulfite chemistry should be the same for nearly all kits. Some offers additional detergents (i.e. Qiagens 'DNA Protection Buffer'). Some kits provides denaturation solutions (sodium hydroxide) while other kits use a thermal denaturation. Incubation temperatures and times differs. Subsequent purification is realized mostly by DNA binding on silica membranes. This methods bases on the same principles and the only parameters with a high influence on downstream applications are from my point of view residual ethanol and sulfite. Both are strong PCR inhibitors.