Patent ID: 9012183
Filing Date: 2015-04-21
Classification: C12N,C12P,C12Q

Abstract:
1. A method of preparing a DNA copy of a target polynucleotide using template switching, comprising: mixing at least one double stranded template/primer substrate, consisting of a DNA primer oligonucleotide annealed with a complementary oligonucleotide template strand, with at least one target polynucleotide in a reaction medium, and adding a suitable amount of a non-retroviral reverse transcriptase to the reaction medium to extend the DNA primer oligonucleotide from its 3′ end to provide a DNA copy polynucleotide that includes a complementary target DNA polynucleotide that is synthesized using the target polynucleotide as a template wherein the double stranded template/primer substrate has a blunt end wherein the 3′ end of the DNA primer oligonucleotide is directly aligned with the 5′ end of the complementary oligonucleotide template strand, or an overhanging end wherein the 3′ end of the DNA primer oligonucleotide extends 1 nucleotide beyond the 5′ end of the complementary oligonucleotide template strand.