The SELEX procedure involves the selection of ligand RNAs from a
library generated from a population of DNAs with randomized sequences.
Amplification of effective ligand RNAs requires reverse transcription of
the RNA pool and subsequent PCR on the cDNA with specific flanking
oligos. Has anyone out there reported the use of ssDNA for the same
purpose? This would negate the need to reverse transcribe and
re-transcribe each time. If one included dUTP in the PCR reaction, then
the products would be able to form G:U base pairs, which may aid the
structural diversity of the pool. Assymetric PCR could be used to
generate ssDNA, or the pool could simply be heated and rapidly cooled to
favor intra-molecular base-pairing. Any references to this would be helpful.
THanks in advance.....
John Weiland jweiland at badlands.nodak.edu