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AssigneeBoard of Regents, The University of Texas System
InventorsAlan M. Lambowitz, Sabine Mohr, Travis B. White, Scott Kuersten
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A method of preparing a DNA copy of a target polynucleotide using template switching is described. The method includes mixing a double stranded template/primer substrate made up of a DNA primer oligonucleotide associated with a complementary oligonucleotide template strand with a 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. The DNA copy polynucleotide includes a complementary target DNA polynucleotide that is synthesized using the target polynucleotide as a template. Methods of adding nucleotides to the double stranded template/primer substrate are also described. The method can be used to facilitate detection, PCR amplification, cloning, and determination of RNA and DNA sequences.
A method of preparing a DNA copy of a target polynucleotide using template switching, comprising:
A method of preparing a DNA copy of a target polynucleotide using template switching, comprising:
A method of adding additional nucleotides to a DNA primer oligonucleotide, comprising adding a suitable amount of a non-retroviral reverse transcriptase to a reaction medium comprising a double stranded template/primer substrate consisting of a DNA primer oligonucleotide associated with a complementary oligonucleotide template strand, wherein the non-retroviral reverse transcriptase adds 1-15 additional non-complementary nucleotides at the 3′ end of the DNA primer oligonucleotide.
Dependencies are inferred from the claim text.