DNA recovery by phenol/chloroform extraction or GeneClean
dunman at njmsa.UMDNJ.EDU
Sat Feb 15 14:18:33 EST 1997
goldberg at bms.com (Steven Goldberg) writes:
>In article <Pine.GSO.3.95L.970209222612.22253B-100000 at uststf2>, Wilson
><netson at uxmail.ust.hk> wrote:
>> Hello everybody,
>> I would like to know which method can recovery DNA from agarose
>> gel is better: Phenol/chloroform extraction or GeneClean? Which one can
>> have higher yield of recovery and purity of DNA?
>> Should I vortex the mixture during phenol/chloroform since I have
>> heard that if the sticky ends DNA was vortexed, they will screw up
>> together. What is the solution to this problem if DNA screw up? Can I
>> solve it by incubation of the DNA at 65C for 15 min?
>After lots of problems using GeneCleaned DNA in ligations, I switched to
>the QIAquick gel extraction kit and have had very good luck in terms of
>yield and purity.
I too have used QIAquick gel extraction kit with very good results. I've
not ran into ligation problems. However, occassionally the kit denatures
small fragments so check your extracted product on a gel. If denaturing
occurs simply reanneal the fragments.
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