Lj. Mou et al., IMPROVEMENTS TO THE DIFFERENTIAL DISPLAY METHOD FOR GENE ANALYSIS, Biochemical and biophysical research communications, 199(2), 1994, pp. 564-569
Differential display requires two primers to identify discordantly exp
ressed mRNAs. Anchored primers with at least one G residue were superi
or to those which had one C residue and those ending in A or T were th
e least efficient. Arbitrary primers with GC pairs at the 5'end were s
uperior to primers with GC pairs at the 3'end. We found that screening
of amplified DNA-fixed membranes by a slot blot manifold and hybridiz
ation with DNA probes made from sample RNA rapidly re-screened with DN
A fragments for differential expression, requires less RNA and is fast
er than Northern analysis. Similarly, hybridization of DNA fragments t
o plasmid DNA fixed to membranes and direct PCR sequencing rapidly det
ermine homology to common laboratory plasmids prior to any further man
ipulation. These modifications permit rapid isolation and characteriza
tion of DNA fragments identified by differential display. (C) 1994 Aca
demic Press, Inc.