Methods for making nucleic acids

US6114152A · kind A · utility

Assignee

• The Regents of the University of California · US

Inventors

• Tito Serafini · San Francisco, US
• Percy Luu · Oakland, US
• John Ngai · Boxborough, US
• David M. Lin · Ithaca, US

Key dates

• Filing date: Mar 27, 1998
• Grant date: Sep 5, 2000
• Expiry date: Mar 27, 2018

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6853
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

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.