PCR amplification before bisulfite conversion
Posted 06 October 2010 - 02:04 PM
I have been working with DNA extracted from paraffin-embedded tissue and I want to do a methylation analysis of a promoter. The problem is that my genomic DNA is not as good as I would like so after bisulfite conversion (Qiagen EpiTech kit), when I try to do a BSP, I canít get any band.
Because of that I am thinking on doing the gene amplification before making the bisulfite conversion. Does anyone know if it is possible? Have anyone try it? Iím not sure if itís going to work because bisulfite conversion degrades the most part of DNA so itís more likely to do it with small DNA fragments. Any ideas?
Posted 06 October 2010 - 02:11 PM
Posted 07 October 2010 - 12:22 AM
Posted 07 October 2010 - 02:34 AM
Posted 07 October 2010 - 05:30 AM
and another question is if I try to amplify the same promotor of a cell line instead of FFPE samples how big is the amplicon I can amplify?? can I try to amplify maybe 1000bp?
Posted 13 October 2010 - 01:31 AM