This invention provides a method for producing a temporally paced
subtracted cDNA library comprising: a) isolating temporally spaced RNAs
from cells; b) generating cDNA inserts from the RNAs isolated from step
(a); c) producing a temporally spaced cDNA library having clones
containing the cDNA inserts generated from step (b); d) producing double
stranded cDNA inserts from the temporally spaced cDNA library; e)
denaturing the double stranded cDNA inserts; f)contacting the denatured
double stranded cDNA inserts produced in step (e) with single-stranded
DNAs from another cDNA library under conditions permitting hybridization
of the single-stranded DNAs and the double-stranded cDNA inserts; g)
separating the hybridized cDNA inserts from the unhybridized inserts; h)
generating a cDNA library of the unhybridized inserts, thereby generating
a temporally spaced subtracted cDNA library.