dNTPs from cells

[Mega [Andreas Stuermer]]

Hi guys,

For what I know TAQ polymerase and T4 Ligase is pretty stable.

dNTPs not so much, although alledgedly without any freeze thaw cycle they retain 90% activity after 7 weeks of storage at room temperature.

Now it should be possible to take a crude bacterial lysate (must be DNAse free) instead of dNTPs. Will look into small molecule extraction protocols tomorrow.

Anyone ever heard of this?

[Mega [Andreas Stuermer]]

"A procedure for rapid extraction and high-pressure liquid chromatographic separation of the nucleotides and other small molecules from bacterial cells" 

They use forimc acid to extract the small molecules. 

However, there is a lot of dNDPs and dNMPs as well as NTPs, NDPs and NMPs present. Maybe high ammounts of AMP inhibits polymerase by indicating that there's little energy left