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Quantification of mRNA on Northern Blots

Joe Horecka horecka at molbio.uoregon.edu
Wed Nov 22 13:11:34 EST 1995


In article <48pfa4$fiq at mserv1.dl.ac.uk>, TOMASKA at fns.uniba.sk wrote:

> Dear Netters,
>     My colleage is studying the effect of heat-shock on
> transcription of her gene. As a standart for quantification of mRNA
> on membrane she tried to use mRNA for actin. Unfortunately, the
> activity of actin gene seems to be also affected by the heat
> treatment. Can somebody suggest:
> 
>     1. Another acceptable standart mRNA?
>     2. A possible source of a probe?
> 
>     Thank you in advance for all suggestions.
> 
>     Lubo Tomaska            tomaska at fns.uniba.sk
>     Department of Genetics
>     Comenius University
>     Faculty of Natural Sciences
>     Mlynska Dolina B-1
>     842 15 Bratislava
>     Slovakia

Lubo,

   I'm sending you DNA to probe for LYS7 mRNA. LYS7 seems to be highly
transcribed and so we get nice results with Northern blots and short film 
exposures. We've found LYS7 to be constitutively transcribed under
conditions of heat-shock, cold-shock, and tunicamycin treatment. Also,
it's not under general amino acid control (published in Gene,1995, vol
162, pp 87-92). 

Regards,

Joe Horecka
horecka at molbio.uoregon.edu



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