Differential Display PCR

Becky Bergstrom Rebecca_Bergstrom at qms1.uiuc.edu
Thu May 27 10:47:35 EST 1993


In article <1993May25.105731.8471 at mbcf.stjude.org>, moore at mbcf.stjude.org
wrote:
> 
> 
> I have been using differential display PCR to determine transcriptional
> differences between two closely related cell types.  This has been thoroughly
> described in Science Vol. 257 Pages 967-971.  Although this has worked fairly
> well I am still expreriencing a few problems and have not found a band of
> interest.  Has anyone out there been using this technique and with what
> success?  Is the efficiency of this procedure as good as stated in the paper?
> Any information would be appreciated.
> 
> 
> -- 
> ============================================================================
> Steve Moore, moore at mbcf.stjude.org
> Dept Immunology
> St. Jude Children's Research Hospital
> Memphis, TN  38101
> 901-522-0478
> =============================================================================
>   I have been using differential display for about 4 months now and I have had some success, I think. I am working on regenerating and nonregenerating tissue differences. We used the RNA map kit from the Gene Hunter Corporation, in MA, to get us started. Since then we have developed our own technique for our particular samples. I get differentially displayed bands with each sample I run usually. I've noticed it's really important to run ladder next to your samples to keep track of their size when clonin


g them into vector. Selected bands should come from the upper 1/3 of the gel. We have been ignoring the stutter bands at the bottom of the gel. What exactly are your problems-getting a differentially displayed band from your two different samples or getting your band cloned? I have problems getting the band cloned in and to cut out at the original size.                                                                             
Hope this gives you some clues. 



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