Methods and primer sets for high throughput PCR sequencing

US10894983B2 · kind B2 · utility

Assignee

• HS DIAGNOMICS GMBH · DE

Inventors

• Julia-Marie Ritter · Berlin, DE
• Volkhard Seitz · Berlin, DE
• Steffen HENNIG · Berlin, DE
• Michael Hummel · Berlin, DE

Key dates

• Filing date: Dec 20, 2013
• Grant date: Jan 19, 2021
• Expiry date: Dec 22, 2034

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6869
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Described herein is a method for amplifying a target nucleic acid sequence ta-tC-tV-tC′-tn′ comprising a first amplification using a first primer pair with sequence ma-K-pC and ma-K′-pC′, and a subsequent second amplification using a second primer pair with sequence aL-aP-aK and aL′-aP′-aK′, wherein pC is the same sequence as sequence element tC. pC and pC′ are 8 to 40 nucleotides in length, K comprises a 3′-terminal sequence k1-k2-s, s is a mismatch sequences preventing PCR bias, ak is the same sequence as sequence element k1, aP-aK hybridize to a contiguous sequence on sequence element ma-K, k1 is 2 to 9 nucleotides in length, aL and aL′ can be any sequence, and tV is a variable region within the target nucleic acid sequence. Also described are collections of primer sets for use in the method of the invention.