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Differential Display Analysis

EBERHARDT at RCF.MAYO.EDU EBERHARDT at RCF.MAYO.EDU
Fri Dec 3 09:38:44 EST 1993


|>From:	FERMAT::"BIOSCI-REQUEST at net.bio.net"    1-DEC-1993 17:43:02.75
|>To:	bio-soft at net.bio.net
|>CC:	
|>Subj:	Differential Display Analysis
|>
|>
|>Jim Owens wrote -
|>
|>> Recently some coworkers have been sequencing some cDNAs derived from
|>> differential display analysis so they are likely to come up with some
|>> unknown sequences.
|>
|>Can somebody post an explanation of what the term "differential display 
|>analysis" means, or better yet a published reference where it's described?
|>
|>Thanks very much.
|>
|>Dave Merberg

	DDPCR is a method developed by Arthur Pardee's laboratory to display 
differences in the populations of two mRNA populations, e.g., cells treated 
with different effectors or normal and tumor cell lines, to visualize and 
enable the cloning of the differentially expressed genes.  It utilizes a 
random set of 20-26 5' primers of unique but differing sequence and a set of 
3' primers of the form NVTTTTTTTTTTT to anchor this primer at the 3' end of 
the polyA tail.  The products are relatively short (100-300 bp) that are 
displayed on sequencing gels.  The difference products can be recovered from 
the gels, reamplified and cloned, providing a unique 3' probe that can be 
compared with the data bases and used to clone a full lenth cDNA from a cDNA 
library.  References are cited below:

1.  Liang and Pardee, Science 257: 967-971 (1992).
2.  Liang et al. Nucl Acids Res 21: 3269-3275 (1993).
3.  Bauer et al. Nucl Acids Res 21: 4272-4280 (1993).

	A related technique has been devised by Michael Wigler's group to 
use PCR to clone the differences between two complex genomes.  The reference 
is Lisitsyn et al. Science 259: 946-951.

Hope this helps.

Norman L. Eberhardt, Ph.D.
Associate Professor
Departments of Medicine and:
Biochemistry/Molecular Biology
4-407 Alfred
Mayo Clinic
Rochester, MN 55905
eberhardt at mayo.edu	





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