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Title: Enhanced DNA Typing Kits for Challenging Forensic DNA Samples
Keywords: Short Tandem Repeats, Forensic DNA Typing, PCR Inhibtion, Degradation, Amelogenin, DNA Profiling
Issue Date: 24-Jun-2010
Citation: LIM ENG SENG, SIMON (2010-06-24). Enhanced DNA Typing Kits for Challenging Forensic DNA Samples. ScholarBank@NUS Repository.
Abstract: Degradation in forensic DNA samples, reliable gender determination and inhibition of the polymerase chain reaction (PCR) process are the main challenges to DNA typing. Using a combination of a Taq mutant polymerase (OmniTaq), EzWayTM PCR Direct Buffer, panel of gender determining markers and reduced-size Short Tandem Repeat (STR) primer sets, developmental validation using Scientific Working Group on DNA Analysis Methods (SWGDAM) guidelines were tested on two miniplexes. Miniplex 1 comprises of the larger STR loci in the AmpFlSTR® Identifiler® PCR Amplification kit (D2S1338, D21S11, CSF1PO, D7S820, D13S317, TPOX, D18S51, D16S539 and FGA) and three gender markers: sex-determining region Y (SRY), Amelogenin and DYS392. Miniplex 2 comprises of the remaining STR loci (TH01, D19S433, D13S317, D3S1358, D2S1776, D5S818, vWA, D8S1179) and two additional STR markers D2S1776 and DYS390. Our results demonstrate that the two miniplexes are highly robust in overcoming PCR inhibitors, provide accurate gender determination and useful in the analysis of degraded DNA. A novel method of a single amplification/detection of Miniplex 1 and Miniplex 2 in a single PCR is also presented.
Appears in Collections:Master's Theses (Open)

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