Patent ID: 6083727
Filing Date: 2000-07-04
Classification: C12N,C12Q

Abstract:
A method of producing double-stranded deoxyribonucleic acid molecules from a single-stranded ribonucleic acid molecule comprising a polyadenylated tail, said method comprising:(a) contacting said ribonucleic acid molecule with an oligo dT primer under conditions sufficient for template driven enzymatic deoxyribonucleic acid synthesis to occur, whereby a first hybrid molecule comprising said ribonucleic acid molecule hybridized to a first deoxyribonucleic acid molecule comprising said oligo dT primer is produced;(b) isolating said first hybrid molecule;(c) contacting said isolated first hybrid molecule with a random primer under conditions sufficient for template driven enzymatic deoxyribonucleic acid synthesis to occur, whereby said first hybrid molecule is converted to a second hybrid molecule comprising said first hybrid molecule and a second deoxyribonucleic acid molecule comprising said random primer and a sequence complementary to the 5' end of said ribonucleic acid molecule; and(d) converting said second hybrid molecule to a first double-stranded deoxyribonucleic acid molecule comprising said oligo dT primer and a second double-stranded deoxyribonucleic acid molecule comprising said random primer.