Extract DNA from gel - (Mar/30/2007 )
Hi, can you help me on that, I tried to Isolate the DNA from the band on 1% agarose (TAE buffer), I used 0.1 V 3M Sodium acetate anhd+ glecial acetic acid = Ph 5.2, with 3 V 70% EThOH, Centrifugation, saw the DNA pellete very tinny, after running the gel, I can't see any band.
I need a short quick method.
Thanks,
Roka
When using a method that does not use column purification it can be difficult. You'll want to cut the most amount of gel away from your band, then dissolve the agarose completely. I would then precipitate the DNA with as close to 100% EtOH as you can get. You will obviously not be able to get to 100%. Wash you pellet with 70%.
the key here is to have enough starting DNA in your band. You're going to lose a lot. How uch did you run in the band?
the key here is to have enough starting DNA in your band. You're going to lose a lot. How uch did you run in the band?
I used 6ul of DNA , Also I tried 12 ul DNA+2ul loading buffer.
Acctualy I tried several methods to get the DNA out of the gel.
1- Using the tinny halled tube .2 ml into normal 1.5ml epend. get the solution out by centrifugation for 10 min.
2- re-melt the agarose by bathwater at least 6 hr. at 40C.
After that I use the method that I mentioned before.
Is the NA acetate enhd+ gelecial acetic acid is ok or can I use different chemicals.
Roka
What I meant was how much DNA (in ugs or ngs) did you load to the gel. You're reagents and methods seem OK, I don't think you're starting with enough DNA in the gel.
Maybe you also have a look at this topic?
K.
K.
Thaks a lot for your help.
Roka