Differential display of rt-pcr products: a tool for identifying
novel gene expression during lung development, injury, and
tumorigenesis.
Sunday, Mary E.
Department of Pathology, Brigham & Women's Hospital and Harvard
Medical School, 75 Francis Street, Boston, MA 02115
APStracts 2:0093L, 1995.
The unique identity of each cell is the result of differential gene
expression. A new strategy for differential cDNA screening introduced
by Liang and Pardee utilize s anchored oligo-dT primers and random 5'
oligonucleotide 10-mers to carry out PCR on reverse-transcribed RNA
from different cell populations. The amplified cDNAs are displayed on
a standard sequencing gel as 100-500 bands per lane on the resulting
autoradiograms and comparisons drawn between the different cell
populations. The major advantages of this method over previous
differential screening approaches are: its high sensitivity, the
small amounts of tissue required, the ability to carry out rapid mRNA
analyses using total RNA and to test multiple tissues in parallel.
Its limitations are: the need for many primer combinations for
adequate representation of mRNAs and the large number of bands
displayed. A screening strategy should include multiple positive and
negative control samples and Northern blots to identify cDNAs
differentially expressed. This approach should facilitate the
identification of many novel genes expressed in a variety of
physiological and pathological conditions.
Received 8 May 1995; accepted in final form 19 May 1995.
APS Manuscript Number L156-5.
Article publication pending Am. J. Physiol. (Lung Cell. Mol.
Physiology).
ISSN 1080-4757 Copyright 1995 The American Physiological Society.
Published in APStracts on 30 May 1995.