University of California

Percy Luu

David W Lin

Tito A Serafini

John J Ngai

Nucleic acids are made by converting a primed single-stranded DNA to a double-stranded DNA by a method comprising the step contacting the single-stranded DNA with a DNA polymerase having 5&prime; exonuclease activity under conditions whereby the DNA polymerase converts the-single stranded DNA to the double-stranded DNA, wherein the single-stranded DNA is primed with oligonucleotide primer comprising a sequence complementary to the 3&prime; end of the single-stranded DNA, and at least one of the 5&prime; end of the primer and the single-stranded DNA comprises an RNA polymerase promoter joined to an upstream (5&prime;) flanking moiety which protects the promoter from the 5&prime; exonuclease activity of the DNA polymerase.

Methods for making nucleic acids

Nucleic acids are made by adding a known nucleotide sequence to the 3' end of a first RNA having a known sequence at the 5' end to form a second RNA and reverse transcribing the second RNA to form a cDNA. In one embodiment, the first RNA is an amplified mRNA, the known sequence at the 5' end comprises a poly(T) sequence, the adding step comprises using a polyadenyltransferase to add a poly(A) sequence to the 3' end, the reverse transcribing step is initiated at a duplex region comprising the poly(T) sequence hybridized to the poly(A) sequence, the cDNA is converted to double-stranded cDNA by a polymerase initiating from a noncovalently joined duplex region, and the double-stranded cDNA is transcribed to form one or more third RNAs.

Growing community of inventors

Oakland, CA, United States of America

Percy Luu