Sunday 29 May 2016

biochemistry - For how long and how cold should I perform ethanol/isopropanol precipitations of RNA or DNA?

I always did mine at -80 C, but I never compared the results to other protocols (I don't fix what's not broken). But, I was curious about the same thing, so I looked around. I found one paper discussing this: Paithankar and Prasad, Nuc Acids Res 19(6):1346 (1991)



It shows that at low concentrations, EtOH at RT actually outperforms the precipitations at both 4 C and dry ice/ethanol bath. That difference is quite big at 100ng/ml DNA and lost when there is more than 10 ug/ml DNA. For typical extractions, based on this data, I'd do it at RT.



On the other hand, Hilario and Mackay say in their book that:




for genomic DNA isolation, different DNA precipitation temperatures
and incubation times have little effect on recovery rates. One can
directly centrifuge after adding ethanol without the -20 C incubation,
and, it -20 C ethanol is not available, room temperature ethanol can
be used.




They do not, however, provide any citation for that statement.

No comments:

Post a Comment