Genomic DNA sample preparation

[Smriti]

Hi There,

I am new to handling genomic DNA so have certain experimental doubts:

1. When we prepare certain dilution of genomic DNA, how do we make sure that genomic DNA solution is homogenously mixed. Is gentle pipetting enough or we should heat the sample at aroung 40-45 C (104-113 F)?

2. What is the procedure of reconsituting powdered form of genomic DNA, can we add buffer and heat it at 65C (149F) to dissolve it properly?

Early response would help me a lot.

Thanks!

[zsmith]

Lyophilized DNA should go back into solution readily in water, Tris, or TE buffer. Generally the more concentrated the DNA is the more difficult it will be to get it to go into solution without heat. 45C should be plenty.

[Smriti]

Thanks zsmith for your reply. So that means just before shearing your sample, you heat every sample to 45C? And after shearing, just before analyzing also, do you heat it again?

Thanks again!