DNA extraction with Phenol:Chloroform - DNA extraction (Jun/13/2007 )
I am hoping I may be able to get some hints as to what to do about DNA I have extracted using Phenol:Chloroform. After adding the Salt solution to percipitate the DNA out and rinsing with EtOH, the mixture is very viscous, but I am unable to get a pellet of DNA seperated from the EtOH and the mixture is not viscous enough to spool. I attempted to pour and seperate, but the entire solution is viscous. Does anyone have any idea why it would do that or suggestions on how to fix it?
the DNA may be too much concentrated. But my best guess is that RNA remains in your preparation. So resuspend all in great volume (and don't hesitate to heat to 65°C) and go for RNase/proteinaseK/phenol chlorofom
fred why you need to use proteinase K??? i dont use proteinase K for extracting DNA by phOH:chcl3
abenson25, what you are extracting?? is it genomic DNA or cDNA??
Proteinase K is useful if extracting genomic DNA isnt it? to get rid of any cell debris.
I use PK in all my DNA purification.