starting amount of DNA - (Nov/05/2008 )

hi
my methylation protocol requires about 5ug of DNA,but the dna i extract is just around 300ng/ul.how does everyone increase or enhance the DNA to such high starting levels.is IVT relevant here.
Thanks

Could you provide more details?

e.g. what are you extracting from? can you just get more cells for example?
Also, you quoted that you get 300ng/ul. What is the total amount that you're getting? Are you even close to 5ug? Also, is this a quantity issue, or a concentration issue?

In any case, if you're just wanting to increase the quantity, maybe try looking into various amplification protocols (e.g. whole genome amp, WGA, MDA etc.)

It depends on what you are going to do with the DNA and what method to use. For bisulfite treatment, 300 ng is good enough. Using too much DNA may actually have the problem of incomplete conversion. I have successfully modified and PCRed DNA from a few cells.

Whole genome amplification probably is not good for DNA methylation study because after the amplification, all methylation is lost.

standard ethanol precipitation and resuspending in a much lower volume to increase the concentration.