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
0
1 reply to this topic
#2
perneseblue
-
- Global Moderators
-
- 530 posts
Unlimited ligation works!
8
Neutral
Neutral
Posted 24 February 2009 - 02:08 AM
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
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.
May your PCR products be long, your protocols short and your boss on holiday
