›› 2014, Vol. 30 ›› Issue (5): 342-345.DOI: 10.3969/j.issn.1004-5619.2014.05.006

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Species Identification and Absolute Quantification of Biological Samples by Droplet Digital PCR

HU WEI, CHEN RONG-HUA, ZHANG CHEN, AN ZHI-YUAN, WANG BING, PING YUAN   

  1. (Shanghai Key Laboratory of Crime Scene Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai 200083, China)
  • Online:2014-10-25 Published:2014-10-28

Abstract: Objective To test droplet digital PCR for species identification and absolute quantification of biological sample. Methods Specific primers and probes for human mtDNA encoding gene ND4 and 16S rRNA were designed, and the species-specificity was assessed on DNA samples derived from human and common animals. To determine the sensitivity and stability of droplet digital PCR for species identification and absolute quantification, gradient dilution series of recombinant plasmid and 16 human DNA samples were analyzed. Results Human recombinant plasmid FAM (ND4) could be used in detecting the samples of human. And the results of detecting were consistent with all levels of diluted concentrations. Droplet digital PCR was able to detect low and single copy of target DNA. Conclusion Droplet digital PCR, with high sensitivity and specificity, is fully amenable for species identification and absolute quantification of biological samples, also it can be applied on routine forensic examination.

Key words: forensic genetics, polymerase chain reaction, species identification, absolute quantification, droplet digital PCR

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