AmpFLSTR™ Identifiler™ Plus PCR Amplification Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Extract total genomic DNA was using the PureLink® Genomic DNA Mini Kit (K1820-02, Invitrogen)	Follow the manufacturer’s instructions	For data analysis: analyze the STR data using the DSMZ (German Collection of Microorganisms and Cell Cultures) online STR database

Upstream tips
Extract total genomic DNA was using the PureLink® Genomic DNA Mini Kit (K1820-02, Invitrogen)
Protocol tips
Follow the manufacturer’s instructions
Downstream tips
For data analysis: analyze the STR data using the DSMZ (German Collection of Microorganisms and Cell Cultures) online STR database

Publication protocol

STR profiling: STR loci and the amelogenin sex-determining marker were amplified using the AmpFlSTR™ Identifiler® Plus PCR Amplification Kit (Applied Biosystems, USA) (detecting amelogenin, D5S818, D21S11, D7S820, CSF1PO, D2S1338, D3S1358, vWA, D8S1179, D16S539, TPOX, TH01, D19S433, D18S51, FGA and D13S317) or the DNA TyperTM15 plus direct kit (Institute of Forensic Science, Ministry of Public Security, China)(detecting the loci listed above plus Penta E, D6S1043 and D12S391) according to the manufacturer’s instructions in a GeneAmp®PCR system 9700. Electrophoretic analysis was performed using a 3730/3130xl DNA Analyzer (Applied Biosystems, USA). After electrophoresis, the data were analyzed with Gene Mapper® ID-X Software v1.2 (Applied Biosystems, USA) to categorize peaks by size in relation to an internal standard allelic ladder.
Data analysis: STR data were analyzed using the DSMZ (German Collection of Microorganisms and Cell Cultures) online STR database (http://www.dsmz.de/fp/cgi-bin/str.html), which includes STR data sets for 2,455 human cell lines from ATCC, DSMZ, JCRB and RIKEN. After registration, our STR data were entered for matching with the authenticated cell lines listed in the database44. At the same time, STR profiles of cell lines were compared to additional reference where available, such as that of donor tissue. Furthermore, all cell lines were checked against database of cross-contaminated or misidentified cell lines on ICLAC (International Cell Line Authentication Committee) (Version 8.0, http://iclac.org/databases/cross-contaminations/).

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