Methods and compositions for rapid multiplex amplification of STR loci

• US 9,310,304 B2
• Filed: 05/11/2012
• Issued: 04/12/2016
• Est. Priority Date: 05/12/2011
• Status: Active Grant

• Current Assignee: ANDE Corp.
• Original Assignee: NetBio, Inc.
• Inventors: Schumm, James W., Selden, Richard F., Tan, Eugene
• Primary Examiner(s): Horlick, Kenneth
• Application Number:
  US13/469,971
• Publication Number:
  US 20120309637A1
• Time in Patent Office: 1,432 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 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence

Abstract

Provided are methods for multiplex polymerase chain reaction (PCR) amplification of short tandem repeat (STR) loci that can be used to rapidly generate a highly specific STR profile from target nucleic acids. The resulting STR profiles are useful for human identification purposes in law enforcement, homeland security, military, intelligence, and paternity testing applications.

Claims

1. A method to detect the presence of single nucleotide polymorphisms (SNPs) of a nucleic acid in a sample containing the nucleic acid, comprising:

(a) contacting in a solution said sample with at least eight unlabeled oligonucleotides under conditions permitting hybridization between said oligonucleotides and the nucleic acids in said sample;

(b) generating primer extension products, each product labeled with one of at least eight different fluorescent dyes by primer extension reactions in the presence of ddNTPs, each ddNTP labeled with one of at least eight different fluorescent dyes; and

(c) detecting the SNPs by detecting the primer extension products, each product labeled with one of at least eight different fluorescent dyes by laser-induced fluorescence using a galvanometer in a step and stare mode and a spectrograph to detect the fluorescence from each of said at least eight different fluorescent dyes.

2. A method to detect the presence of single nucleotide polymorphisms (SNPs) of a nucleic acid in a sample containing the nucleic acid, comprising:

(a) contacting in a solution said sample with at least eight oligonucleotides, each labeled with a different fluorescent dye, under conditions permitting hybridization between said oligonucleotides and the nucleic acid in said sample; and

(b) detecting the SNPs by detecting the hybridization products, each product labeled with one of at least eight different fluorescent dyes by laser-induced fluorescence using a galvanometer in a step and stare mode and a spectrograph to detect the fluorescence from each of said at least eight different fluorescent dyes.