Methods and reagents for reducing non-specific amplification

• US 9,410,195 B2
• Filed: 07/17/2015
• Issued: 08/09/2016
• Est. Priority Date: 12/22/2011
• Status: Active Grant

• Current Assignee: Roche Molecular Systems (Roche Holding AG)
• Original Assignee: Roche Molecular Systems (Roche Holding AG)
• Inventors: Bodepudi, Veeraiah, Schoenbrunner, Nancy, Will, Stephen
• Primary Examiner(s): THOMAS, DAVID C
• Application Number:
  US14/802,481
• Publication Number:
  US 20150322502A1
• Time in Patent Office: 389 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12Q 1/6848 characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2525/117 incorporating modified base

Abstract

The present invention provides reagents for use in the amplification of nucleic acids. Amplification carried out using oligonucleotides containing modified nucleotides can result in less non-specific amplification compared to amplification carried out using unmodified oligonucleotides.

Claims

1. A reaction mixture for the amplification of three or more target nucleic acids, comprising three or more pairs of primer oligonucleotides and three or more probe oligonucleotides wherein at least one probe oligonucleotide incorporates a N2-benzyl-dG nucleotide.

2. The reaction mixture of claim 1 further comprising a nucleotide-incorporating biocatalyst, nucleoside triphosphates, and a buffer suitable for the amplification of said three or more target nucleic acids by the nucleotide-incorporating biocatalyst.

3. A kit for the amplification of three or more target nucleic acids, comprising three or more pairs of primer oligonucleotides, and three or more probe oligonucleotides wherein at least one probe oligonucleotide incorporates a N2-benzyl-dG nucleotide, at least one nucleotide-incorporating biocatalyst, nucleoside triphosphates, a buffer suitable for the amplification of said three or more target nucleic acids by the at least one nucleotide-incorporating biocatalyst, and a set of instructions for performing the amplification of nucleic acids.