Hi all!
am gonna to study the methylation status for target promoter gene . I have seen that there are alot of kits can be used such as Zymo kits and Qiagen kits but which one is the best ,also in each one there are different kits . So which one exactly is best for me for studying the DNA from blood samples ?
Also if you kow there is an OPTI-Bisulfite method ? Any one used it ?
Saurabh Mandal which one of the zymo kits ? I have seen 4 types of them (Direct, Gold,lightning and classical) if you know ?
EZ DNA Methylation-GoldTM Kit, this is the product I noticed many researchers are using it. Even our lab is using the same. You can check these papers.
Article Evaluation of bisulfite kits for DNA methylation profiling i...
Article Performance Evaluation of Kits for Bisulfite-Conversion of D...
Article Comparative analysis of 12 different kits for bisulfite conv...
Article Comprehensive Evaluation of Commercial Bisulfite-Based DNA M...
Zymo kits have an excellent reputation. You should also consider the protocol, as it has been shown that longer incubation periods with cycling (e.g. 16 cycles of 95oC for 30s + 50oC for 60mins) achieve the best results:
Article Pyrosequencing Evaluation of Widely Available Bisulfite Conv...
Agree with everyone, I used Zymo's kit myself and never had an issue, really good quality. I used the EZ DNA Methylation-Gold Kit.
Just remember when you are going on to qMSP you will only recover >75% of the imputed DNA so you need to factor that in in some way. I found utilising an internal control gene the best way vs a calibrator that may vary by 25% after conversion. Afterwards I used the formula 2^(ct b actin-ct target)*100
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