methylation specific PCR - new to epigenetics-where to start (Jan/02/2009 )
Hi,
I'm new to epigenetics, have never done it before but i'm keen on trying this out.
Is my approach correct?
1) Initially to check if my gene of interest is epigenetically silenced, I treat the genomic DNA (from cell lines) with 5-azadC.
2) If cell lines show pigentic silencing, eh next step would be to check for promoter metylation by methylation specific PCR?
How can I get the promoter sequence of my gene of interest DKK3?
-Sarwat-
1. No. You should treat cell lines with 5-azadC.
2. Correct.
QUOTE (Sarwat @ Jan 2 2009, 08:53 PM)
Hi,
I'm new to epigenetics, have never done it before but i'm keen on trying this out.
Is my approach correct?
1) Initially to check if my gene of interest is epigenetically silenced, I treat the genomic DNA (from cell lines) with 5-azadC.
2) If cell lines show pigentic silencing, eh next step would be to check for promoter metylation by methylation specific PCR?
How can I get the promoter sequence of my gene of interest DKK3?
I'm new to epigenetics, have never done it before but i'm keen on trying this out.
Is my approach correct?
1) Initially to check if my gene of interest is epigenetically silenced, I treat the genomic DNA (from cell lines) with 5-azadC.
2) If cell lines show pigentic silencing, eh next step would be to check for promoter metylation by methylation specific PCR?
How can I get the promoter sequence of my gene of interest DKK3?
-microlight-