Dissolve my pellet, please!
Christoph.Lorra at urz.uni-heidelberg.de
Christoph.Lorra at urz.uni-heidelberg.de
Thu Feb 6 17:24:13 EST 1997
In <32F7A202.212F at codon.nih.gov>, jberke at CODON.NIH.GOV (Josh Berke) writes:
>Hi:
>
>I have pellets of cDNA that just do NOT want to dissolve.
>
>I performed a phenol:chloroform extraction followed by
>a phenol:chloro:IAA extraction. I precipitated the DNA using
>4M NH40Ac and 95% ethanol, with 20min room temp spin at 14,000.
>This was done in a siliconized 0.5ml tube (from Ambion). After
>removing the supernatant I could see the pellet; the cDNA is also
>radiolabelled so I can keep track of it....
Hi
this sounds indeed a bit difficult.
Because you have vortexed the sample you might have destroyed it anyhow.
If not I would try to wash it again several times with 70 % ethanol and afterwards
try to resuspend it in a bigger volume of TE ( make sure the pH is around 8 ).
A high pH might help because of the salt you used.
Afterwards you can precipitate it again with 1/10 volume 3 M NaAcetat pH 5
and 3 volume EtOH, wash 2 times with 70 % EtOH and disolve in TE pH 7.9 or 8.
Hope this will help
Ciao Christoph
More information about the Methods
mailing list