Powerplex vr 1.2 system

STR DNA profiling was carried out for the MaPac107 primary cell line, after establishment, using fluorescent PCR in combination with capillary electrophoresis, as described previously [103 (link)]. Using different alternate colours, the PowerPlex VR 1.2 system (Promega, Mannheim, Germany) was modified in order to run a two-colour DNA profiling, allowing the simultaneous single-tube amplification of eight polymorphic STR loci and Amelogenin for gender determination. The STR loci of CSF1PO, TPOX, TH01, vWA, and Amelogenin were amplified by primers labelled with the Beckman/Coulter dye D3 (green; Sigma-Aldrich, Taufkirchen, Germany), while the STR loci D16S539, D7S820, D13S317, and D5S818 were amplified using primers labelled with D2 (black). All the loci except the Amelogenin gene in this set are true tetranucleotide repeats. All primers are identical to the PowerPlexVR 1.2 system except the fluorescent colour. Data were analysed with the CEQ 8800 software (Beckman-Coulter, Krefeld, Germany), which enables an automatic assignment of genotypes and automatic export of resulting numeric allele codes into the reference DNA database of the DSMZ.

To eliminate the risk of cross contamination STR DNA profiling was carried out using fluorescent PCR in combination with capillary electrophoresis as described previously [20 (link)]. Briefly, the PowerPlex VR 1.2 system (Promega, Mannheim, Germany) was set to run two-color DNA profiling allowing the simultaneous single-tube amplification of eight polymorphic STR loci plus amelogenin (gender). Loci were amplified by primers labeled with the Beckman/Coulter dye D3 (green; Sigma-Aldrich, Munich/Germany), while the STR loci D16S539, D7S820, D13S317 and D5S818 were amplified using primers labeled with D2 (black). Data (Table A in S1 File) were analyzed with the CEQ 8000 software (Beckman-Coulter, Krefeld, Germany), which enables an automatic assignment of genotypes and automatic export of resulting numeric allele codes into the reference DNA database of the DSMZ.

STR DNA profiling was carried out using fluorescent PCR in combination with capillary electrophoresis as described previously^{28 (link)}. Using different alternate colours, the PowerPlex VR 1.2 system (Promega, Mannheim, Germany) was modified in order to run a two-color DNA profiling allowing the simultaneous single-tube amplification of eight polymorphic STR loci and Amelogenin for gender determination. STR loci of CSF1PO, TPOX, TH01, vWA and Amelogenin were amplified by primers labelled with the Beckman/Coulter dye D3 (green; Sigma-Aldrich, Germany), while the STR loci D16S539, D7S820, D13S317 and D5S818 were amplified using primers labelled with D2 (black). All the loci except the Amelogenin gene in this set are true tetranucleotide repeats. All primers are identical to the PowerPlexVR 1.2 system except the fluorescent colour. Data were analysed with the CEQ 8800 software (Beckman-Coulter, Germany), which enables an automatic assignment of genotypes and automatic export of resulting numeric allele codes into the reference DNA database of the DSMZ.