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For your BSA/milk blocking system, if there is a high background then may be you can try optimize the primary/secondary dilutions. I just had this problem a few weeks ago, so what I did was run a gel of the exact same sample, and then cut off each lane and then try out different combinations of primary and secondary dilutions, and see if it gets better.
#122510 Sox-2 antibody detection using Western
Posted
on 27 October 2011 - 10:41 PM
#115782 effect of DNA isolation and storage on methylation
Posted
on 22 July 2011 - 05:28 AM
genie, on 21 July 2011 - 08:14 PM, said:
Hi 
All,
Im new to methylation studies and hope to learn more about methylation.
i want to know if the isolation procedure of DNA has an impact on methylation of genes?
should i use column purification kits or can i even used standard phenol-chloroform method for DNA isolation?
Also can anyone tell me if storage of the isolated DNA in -20C can alter the actual CpG pattern of the genes?
All,Im new to methylation studies and hope to learn more about methylation.
i want to know if the isolation procedure of DNA has an impact on methylation of genes?
should i use column purification kits or can i even used standard phenol-chloroform method for DNA isolation?
Also can anyone tell me if storage of the isolated DNA in -20C can alter the actual CpG pattern of the genes?
Nothing will change the methylation status of your DNA, neither any isolation procedure nor your storage conditions.
Best,
MoB
