hi mates,
I have to separate and extract DNA from a gel. The problem comes because the insert is 4.5kb and the plasmid 4kb. Do I have to use a .7% or 1% agarose? is it alright to use TAE or is better for this case the TBE?
thanks in advance
-t
0
2 replies to this topic
#2
phage434
-
- Global Moderators
-
- 1,805 posts
Veteran
130
Excellent
Excellent
Posted 01 July 2010 - 05:54 PM
Your best bet would be to find an enzyme that cuts the vector but not the desired insert. Then you would have an easy time with the gel. I would use TAE and a 1% gel.
You might want to run it slowly to reduce heat and diffusion.
You might want to run it slowly to reduce heat and diffusion.
#3
toniX
-
- Members
-
- 2 posts
member
0
Neutral
Neutral
Posted 01 July 2010 - 06:33 PM
phage434, on Jul 1 2010, 06:54 PM, said:
Your best bet would be to find an enzyme that cuts the vector but not the desired insert. Then you would have an easy time with the gel. I would use TAE and a 1% gel.
You might want to run it slowly to reduce heat and diffusion.
You might want to run it slowly to reduce heat and diffusion.
I already did a test and the problem was the separation of the two bands (I have to admit that I used a .7% gel and I ran it like there will not be tomorrow, at 150V). I will follow your indications
-t
