Use of restriction enzyme digestion before bisulfite conversion - What is the purpose of r-enzyme digestion? (Jul/05/2005 )

Hi every body,

I've see in a article(Singh et al) where they digested the DNA with Eco RI for over night at 37 degree centi, before bisulfite conversion of DNA for DNA methylation analysis studies. Iam doing the same sought of work, by using EZ DNA methylation kit for bisulfite conversion of DNA. Iam not doing any restriction enzyme digestion in the process and iam unclear about the purpose of that restriction enzyme digestion. It would be great if any body could explain me about the purpose of digestion of DNA with Eco RI or any rstriction enzyme before bisulfite conversion and make my doubt clear.


Ravi

Restriction digestion is not a must. The purpose is to reduce the complexity of genomic DNA so that it will get denatured easily. Alternative you can use a syringe to shear DNA.

Please check this post
http://www.protocol-online.org/forums/inde...indpost&p=17002

Thanks a lot PCR man

Ravi