in vitro transcriptions

Michael Coady 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.

Mike Coady
COADY at ERE.UMONTREAL.CA



More information about the Methods mailing list