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Multiplex Short Tandem Repeat Amplification of Low Template DNA Samples with the Addition of Proofreading Enzymes *
Author(s) -
Davis Carey P.,
Chelland Lynzee A.,
Pavlova Victoria R.,
Illescas María J.,
Brown Kelly L.,
Cruz Tracey Dawson
Publication year - 2011
Publication title -
journal of forensic sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.715
H-Index - 96
eISSN - 1556-4029
pISSN - 0022-1198
DOI - 10.1111/j.1556-4029.2011.01727.x
Subject(s) - proofreading , dna , multiplex , str analysis , computational biology , multiplex polymerase chain reaction , dna polymerase , microbiology and biotechnology , microsatellite , polymerase , biology , chemistry , polymerase chain reaction , genetics , gene , allele
With <100 pg of template DNA, routine short tandem repeat (STR) analysis often fails, resulting in no or partial profiles and increased stochastic effects. To overcome this, some have investigated preamplification methods that include the addition of proofreading enzymes to the PCR cocktail. This project sought to determine whether adding proofreading polymerases directly in the STR amplification mixture would improve the reaction when little template DNA is available. Platinum Taq High Fidelity and GeneAmp High Fidelity were tested in Profiler Plus™ STR reactions alone and in combination with AmpliTaq ® Gold. All reactions included the additional step of a post‐PCR purification step. With both pristine low template DNA and casework samples, the addition of these polymerases resulted in comparable or no improvement in the STR amplification signal. Further, stochastic effects and artifacts were observed equally across all enzyme conditions. Based on these studies, the addition of these proofreading enzymes to a multiplex STR amplification is not recommended for low template DNA work.