Open AccessA short interspersed nuclear element-based quantitative PCR assay for simultaneous human and dog DNA detection and quantification
Author(s) -
James W Liang,
Heather Miller Coyle
Publication year - 2021
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2020-0144
Subject(s) - biology , dna , alu element , microbiology and biotechnology , polymerase chain reaction , real time polymerase chain reaction , low copy number , computational biology , genetics , gene , human genome , genome , plasmid
This study aimed to develop a quantitative PCR assay to simultaneously quantify human and dog DNA in a multispecies mixture to inform downstream analyses in routine forensic casework and scientific research. The human target is the Alu Yb8 element, which has approximately 2000 copies per cell, and the canine target is the SINEC_Cf element, which has approximately 200,000 copies per cell. The internal positive control is a universal exogenous assay consisting of forward and reverse primers, a NED-labeled probe with an MGBNFQ quencher and a 65-bp synthetic template. Results suggest a potentially robust assay with a fast run time and a high degree of sensitivity, precision and species specificity, with direct application to domestic pet samples in veterinary genetics and forensics.