Purify large fragment from Agarose gel >10 Kb

Michelle Gleeson michelle at MOLECULE.BIO.UTS.EDU.AU
Mon Oct 20 20:54:44 EST 1997


Hi,

I recently had success with a simple home made spin column that was
described here a while back:  0.5ml tube with 22G needle hole in bottom,
small amount siliconised glass wool inside.  Cut off cap, place in 1.5ml
tube. Add gel slice from 1% gel into inner tube.  Spin 10 minutes 6000rpm.
Phe:Chl then Chl extract and EtoH ppt the flow through.  I was able to
successfully ligate a 9kb product using this technique, but a 20kb band
also purified well.

AATAGGCAATGGGCCCCATATAGGAACACAGAGCTGCATGCGTATTGCATGCCAGGCTATTCATTCCAGGGAAA
Michelle Gleeson
Molecular Parasitology Unit              Ph: (02) 9514 4043
University of Technology                 Fax:(02) 9514 4003
Westbourne St Gore Hill, NSW 2065        michelle.gleeson at uts.edu.au
When you get to the end of your rope, tie a knot and hang on - FDR
TTATCCGTTACCCGGGGTATATCCTTGTGTCTCGACGTACGCATAACGTACGGTCCGATAAGTAAGGTCCCTTT

On 20 Oct 1997, Ke-Xue Huang wrote:

> Hi, Netters,
>
> Does anybody out there any give me some references or any methods, kits,
> which work well for purification large fragment from agarose gel?(>10 kB)
>
> We met unstable results by using Geneclean or Qiagen kit.
>
> Thanks!
>
>
> Huang
>
>
>




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