Transcritional Start Site
rhaun at nih.gov
Thu Dec 15 17:33:05 EST 1994
In article <3cpr80$s3j at solaris.cc.vt.edu> klingj at vt.edu (klingj) writes:
>From: klingj at vt.edu (klingj)
>Subject: Re: Transcritional Start Site
>Date: 15 Dec 1994 16:35:12 GMT
>In article <3ci858$hej at solaris.cc.vt.edu>, klingj at vt.edu (klingj) says:
>> I am interested in determining the transcriptional start site of a recently cloned
>> gene. What current methods are available..... Please E-mail me at klingj at vt.edu.
>In a follow up to my first question, several people at VA Tech thought that one can
> use RACE-PCR to identify the transcription start site, but nobody had any info on it.
>Is this a viable alternative. I'd also like to avoid isotopes if it is possible. Please let
> me know if anyone has had experience with this technique and if it can be used for
> this purpose. Thanks.
> Jim Kling
>1400 Prices Fork Rd
>Blacksburg, VA 24061-0305
Traditional methods of mapping a transcription start (e.g., nuclease
protection assays and/or primer extension) would be more convincing.
However, these methods are typically performed using radioactive probes.
5'-RACE may get you close to the 5' end of the mRNA, but it would be
difficult to prove that your longest PCR product corresponds to the
transcription start site rather than just a truncation without using one
of these other methods.
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