Patent ID: 6706481
Filing Date: 2004-03-16
Classification: C12N,C12Q

Abstract:
A method of selecting signaling aptamers in vitro comprising the steps of:(a) synthesizing a DNA pool, said DNA having a random insert of fifty-one nucleotides, wherein a A:C:G:T mole ratio of amounts of each nucleotide in said random insert is skewed such that the amounts of only three of said nucleotides are about equal (b) amplifying the DNA pool; (c) transcribing an RNA pool from said amplified DNA wherein a nucleotide used in the RNA transcription is labeled with one or more reporter molecules; (d) applying the labeled RNA pool to an affinity column wherein labeled RNA molecules having a high binding affinity for the column are retained on the affinity column and thereby removed from the labeled RNA pool; (e) obtaining a cDNA pool from the labeled RNA molecules retained on the affinity column; (f) repeating steps (a) through (e) to retain the labeled RNAs from the labeled RNA pool having a high binding affinity for the column on the affinity column; and (g) cloning the retained labeled RNA molecules, said clones comprising signaling aptamers.