in vitro transcriptions
coady at ERE.UMontreal.CA
Wed Jun 23 08:19:03 EST 1993
In article <C91MwF.3u0 at news.Hawaii.Edu> markh at ahi.pbrc.hawaii.edu (Mark Henteleff) writes:
>I am currently doing in vitro transcription protocols to make
>mRNA for Na channel (approx. 6kb) to use in expression in
>Xenopus oocytes. Anybody out there know if its necessary
>to get rid of unincorporated nucleotides before the sample can
>be used in these kinds of expression experiments?
>Thanks Mark H.
If you phenol/chloroform your sample to remove the proteins and
then precipitate once with ammonium acetate and ethanol, you should remove
the majority of the ribonucleotides. Naturally, rinse with 70% EtOH and
then DRY!! Oocytes really do not like ethanol.
COADY at ERE.UMONTREAL.CA
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