genome DNA doesn't dissolve in NaOH - genome DNA doesn't dissolve in NaOH (Dec/11/2007 )

Dear all,

By using trizol reagent i got a huge genome (Cloudy) pellet from human tissue, however, it seems like the pellet can't be dissolve in NaOH. I need to do PCR in the nest step, so, what should i do?

Thank you in advance for your kindly help

freshman in molecular science

I don't know what type of protocol you are using, but the DNA should be dissolve in 1XTE not in NaOH.

Yeah, dissolve it in TE or DNAse free water. Why the NaOH?

I've actually just recently heard that dissolving in NaOH can be done to degrade RNA, although that said, I have no idea about the concentration you should use (another person in my lab said he thought he had read 50mM, but don't rely on that!!).

Good luck

i have a feeling that your cloudy pellet is heavily contaminated with protein and other junk, thus the cloudiness and lack of solubility.

Try conducting 1 or 2 phenol/chloroform extractions on the DNA.
You could try heating your DNA to help get in into solution.