Prepfiler automated forensic dna extraction system

Genomic DNA was extracted from all samples using the automated platform, Automate Express Forensic DNA Extraction System (Thermo Fisher Scientific Inc., Waltham, USA) with the Prepfiler Automated Forensic DNA Extraction System (Thermo Fisher Scientific Inc.). DNA extracts were quantified using the Quantifiler Trio Human DNA Quantification Kit (Thermo Fisher Scientific Inc.) on a 7500 Real-Time PCR System (Thermo Fisher Scientific Inc.). All procedures followed the manufacturers instructions, with elution volumes of 50 mL. A sample was considered not degraded/inhibited when the degradation index (DI) <1.5 and no IPCCT flag was triggered in Quantifiler Trio assay.

Total DNA was extracted using the automated platform Automate Express Forensic DNA Extraction System (Thermo Fisher Scientific Inc.) with the Prepfiler Automated Forensic DNA Extraction System (Thermo Fisher Scientific Inc.), or manually via the DNA IQ System (Promega Corp., Madison, WI, USA). Autosomal DNA was quantified with the Quantifiler Human DNA Quantification Kit (Thermo Fisher Scientific Inc.) on a 7500 Real-Time PCR System (Thermo Fisher Scientific Inc.). MtDNA quantities were estimated using a 1:100 genomic to mitochondrial DNA copies ratio according to the product manual. All procedures followed the manufacturer´s instructions.

Oral swabs or peripheral blood were obtained from 432 unrelated volunteer donors from all five different Brazilian geopolitical regions. Male contributors were preferred, in order to assure a significant sample size for investigation of SNPs located in the Y chromosome included in this study. Number of samples obtained from each Brazilian region and its States are presented in Supplementary Figure S1. Genomic DNA was extracted using the automated platform Automate Express Forensic DNA Extraction System (Thermo Fischer Scientific Inc.) with the Prepfiler Automated Forensic DNA Extraction System (Thermo Fischer Scientific Inc.), or manually using DNA IQ System (Promega Corp., Madison, WI, USA). DNA was quantified with the Quantifiler Human DNA Quantification Kit (Thermo Fischer Scientific Inc.) on a 7500 Real-Time PCR System (Thermo Fischer Scientific Inc.) or using Qubit 2.0 Fluorometer with Qubit dsDNA HS Assay Kit (Thermo Fischer Scientific Inc.). All procedures followed manu-facturer´s instructions.