why not working msp primers - (May/20/2015 )

Hi

I have 3 genes to study MS PCR  but I tried 5 times msp primer desingn with meth primer programs didn’t get any results. Afterward I designing my own primers without any programs, I tried different temperature and time; again failure.   My times is running out and I dont know what can ı do. These gene promoters are difficult for primer designing. I attach my sequences and my own primers file. Could you please help

 Thank you

Hi zyk, how are you bisulfite converting your DNA? are you running the controls that are usually included in the kit?

cheers

Nick

Hi

I converting DNA with EZ methylation gold kit (ZYMO). The kit not involved controls . Therefore I controlled bisufite converted DNA with MSP primers that I used before and they worked.

Thank you for your reply

I would suggest you revisit primer design and use methprimer express from Thermo Fisher.

Thank you i will try