|>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.
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.
Departments of Medicine and:
Rochester, MN 55905
eberhardt at mayo.edu