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/).
Full paper
Login or
join for free to view the full paper.
Videos
Check out videos that might be relevant for performing Cell line authentication MCF-7 cell line using AmpFLSTR™ Identifiler™ Plus PCR Amplification Kit from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.
We haven't found any additional videos for this experiment / product combination yet.