Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Biology

get rid of RNA from Maxi - (Feb/24/2009 )

Hi,

I need a lot of DNA for the transfection. When I do the maxi prep from 500ml, I meet a problem which there is so much RNA. I tried RNase, after percipitation, which does not help much. Any suggetion is greatly appreciated.

Jihua

-jihuafan-

jihuafan on Feb 24 2009, 01:27 AM said:

Hi,

I need a lot of DNA for the transfection. When I do the maxi prep from 500ml, I meet a problem which there is so much RNA. I tried RNase, after percipitation, which does not help much. Any suggetion is greatly appreciated.

Jihua


use lithium chloride precipitation.

Resuspend your DNA in 10ml of TE,
Add equal volume of 8M LiCl (This will precipitate the RNA).
Invert tube to mix
Leave on ice for 15min (on place in freezer to chill faster)
Spin at down for 15min in centrifuge
Move the supernatant to a new tube. Discard the pellet.
add equal volume of isopropanol to precipitate the DNA.

-perneseblue-