RT-PCR-Multiple bands-unusual problem

Tracy Aquilla aquilla at salus.med.uvm.edu
Wed Jun 26 10:19:45 EST 1996


In Article <31CFD1CE.2C88 at facstaff.wisc.edu>, Paul Mozdziak
<pemozdzi at facstaff.wisc.edu> wrote:
>I'm doing some RT-PCR to see if a particular factor is expressed in 
>muscle. First I am using previously published primers. Second my 
>positive control is a small amount of a cDNA probe for my factor of 
>interest. The probe samples amplify beautifully. However, when I run 
>my rt-pcr stuff from muscle out on a 3% gel (sigma agarose (50-1000 
>bp) I get two bands (one at 150 bp--what I should get and one a few base 
>pairs smaller)--they just barely resolve. I ran these samples out 
>shortly after I finished the pcr. 
>
>Any ideas to explain my problem and too solve it?
>
>My RNA was not DNAsed--and it's just total RNA. There was alot of RNA at 
>the beginning of the procedure in my muscle samples ~1 microgram.
>
>Thanks much
>
>Paul

What gene? Many muscle-specific genes have multiple isoforms, some due to a
second gene, some due to alternative splicing of the mRNA.
Tracy

Thomas T. Aquilla, Ph.D.         .***.   .***.           .***.   .***.
Molecular Physiology and Biophysics    * | | | *       * | | | * | | | *
University of Vermont Medical College*   * | | | *   * | | | *   * | | | *   
Health Science Complex, Given E-213*       * | | | * | | | *       * | | | *
Burlington, VT 05405-0068    '***'           '***'   '***'           '***'



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