Post by djoser-xyyman on Mar 23, 2018 8:45:23 GMT -5
www.promega.com/resources/profiles-in-dna/2016/analyzing-data-from-next-generation-sequencers-using-the-powerseq-automitoy-system/
Introduction
Next-generation sequencing (NGS) is a powerful technology that can provide forensic DNA profiles compatible with current databases, plus deliver additional levels of genetic data that will open new doors to investigations, mixture interpretation, missing persons cases, and more. The earliest published reports on the use of NGS for analysis of forensic markers were as recent as 2011–12 (1) (2) (3) . Since this time, greater than 60 peer-reviewed studies using NGS laboratory methods and/or custom software tools have been reported for forensic examination of short tandem repeats (STRs), mitochondria and single nucleotide polymorphisms (SNPs). A full list can be viewed online at: www.genomicidlab.com/opendata
Promega Corporation developed an autosomal STR kit optimized for downstream sequencing with NGS called PowerSeq® Auto System that amplifies 23 loci, including the full CODIS extended STR panel. Developmental validation studies with PowerSeq® Auto System have been performed, demonstrating backwards compatibility to existing STR databases and extremely high sequence diversity in some CODIS core loci (4) (5) . This sequence data may be used to augment STR analysis in samples for which capillary electrophoresis methods are inconclusive (e.g., degradation and mixtures) or additional information could assist the interpretation and reporting (e.g., familial searching and kinship). PowerSeq® Auto System has undergone further development to include Y-chromosome STRs and the control region of the mitochondria. This multiplex forensic NGS kit is called PowerSeq® Auto/Mito/Y System and features small amplicons (129–303bp), high sensitivity (~100pg DNA) and data for three forensic panels (22 autosomal STRs, 23 Y-STRs and 10 amplicons covering the mitochondrial and amelogenin control region). Herein we report on the evaluation of PowerSeq® Auto/Mito/Y System for analysis of reference samples.
Discussion
The PowerSeq® Auto/Mito/Y System is a powerful new system that can be added to the forensic DNA analysis toolkit to help meet the needs of complex sample analysis. The ability to simultaneously analyze autosomal- and Y-STRs along with mitochondrial data from one sample will likely add value to many forensic casework, databasing, and missing persons laboratories. However, for this technology to be fully realized, some aspects need to be addressed. First, forensic sequence databases must be developed to statistically interpret profiles. We are currently addressing this need through a sequence population database to be presented to the Criminal Justice community late 2016 (NIJ Award 2015-DN-BX-K062). Second, laboratory methods need to be streamlined, optimized and validated in simple, low-cost workflows. Third, the software and analysis tools need to be fully developed and validated for the forensic laboratory. Lastly, NGS standards and guidelines need to be defined by governing bodies to allow implementation in accredited laboratory systems. Once these items are addressed, a system such as the PowerSeq® Auto/Mito/Y System can routinely be utilized by crime laboratories to maximize DNA data output from challenging forensic samples.
Introduction
Next-generation sequencing (NGS) is a powerful technology that can provide forensic DNA profiles compatible with current databases, plus deliver additional levels of genetic data that will open new doors to investigations, mixture interpretation, missing persons cases, and more. The earliest published reports on the use of NGS for analysis of forensic markers were as recent as 2011–12 (1) (2) (3) . Since this time, greater than 60 peer-reviewed studies using NGS laboratory methods and/or custom software tools have been reported for forensic examination of short tandem repeats (STRs), mitochondria and single nucleotide polymorphisms (SNPs). A full list can be viewed online at: www.genomicidlab.com/opendata
Promega Corporation developed an autosomal STR kit optimized for downstream sequencing with NGS called PowerSeq® Auto System that amplifies 23 loci, including the full CODIS extended STR panel. Developmental validation studies with PowerSeq® Auto System have been performed, demonstrating backwards compatibility to existing STR databases and extremely high sequence diversity in some CODIS core loci (4) (5) . This sequence data may be used to augment STR analysis in samples for which capillary electrophoresis methods are inconclusive (e.g., degradation and mixtures) or additional information could assist the interpretation and reporting (e.g., familial searching and kinship). PowerSeq® Auto System has undergone further development to include Y-chromosome STRs and the control region of the mitochondria. This multiplex forensic NGS kit is called PowerSeq® Auto/Mito/Y System and features small amplicons (129–303bp), high sensitivity (~100pg DNA) and data for three forensic panels (22 autosomal STRs, 23 Y-STRs and 10 amplicons covering the mitochondrial and amelogenin control region). Herein we report on the evaluation of PowerSeq® Auto/Mito/Y System for analysis of reference samples.
Discussion
The PowerSeq® Auto/Mito/Y System is a powerful new system that can be added to the forensic DNA analysis toolkit to help meet the needs of complex sample analysis. The ability to simultaneously analyze autosomal- and Y-STRs along with mitochondrial data from one sample will likely add value to many forensic casework, databasing, and missing persons laboratories. However, for this technology to be fully realized, some aspects need to be addressed. First, forensic sequence databases must be developed to statistically interpret profiles. We are currently addressing this need through a sequence population database to be presented to the Criminal Justice community late 2016 (NIJ Award 2015-DN-BX-K062). Second, laboratory methods need to be streamlined, optimized and validated in simple, low-cost workflows. Third, the software and analysis tools need to be fully developed and validated for the forensic laboratory. Lastly, NGS standards and guidelines need to be defined by governing bodies to allow implementation in accredited laboratory systems. Once these items are addressed, a system such as the PowerSeq® Auto/Mito/Y System can routinely be utilized by crime laboratories to maximize DNA data output from challenging forensic samples.