How to get a sufficent DNA from gel for ligation?
SerAln at yahoo.com
Fri Oct 25 07:45:52 EST 2002
I'm unable to read the original message, so maybe my reply is out of the
subject. I've tried several "gene clean" kits and I have to say that I still use
the phenol extraction for purifying DNA from gels. No low-melting agarose nor
any kind of resin. Why?. Because I always get tons of DNA with phenol extraction
compared with any of the resins.
It's more time consuming, but I'm sure that loading one or two micrograms of DNA
I'll be able of recovering almost the whole amount afterwards.
> tevinxxx at hotmail.com wrote:
>>Every time, I cut DNA from the gel, but my ligation product did not show enough growth for transformation. Does anyone have better ideas for gel elution process and ligation?
> I use the Quiagen gel extraction kit. Usually about 100 ng of vector DNA is
> sufficient for one ligation reaction (20 microliters). The kit will give you a
> good yield depending on the starting amout of cut vector that you run on gel.
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