Article Title: Applications of Probe Capture Enrichment Next Generation Sequencing for Whole Mitochondrial Genome and 426 Nuclear SNPs for Forensically Challenging Samples
Figure Lengend Snippet: Novel probe capture next generation sequencing (NGS) assay for sequencing mitochondrial (mt) and nuclear DNA. To circumvent choosing between mitochondrial and nuclear analysis of degraded or limited forensic DNA samples, a capture enrichment based library preparation method can be implemented. In this method: ( 1 ) DNA libraries are prepared by fragmenting the DNA samples using Covaris ® M220 Focused-ultrasonicator TM (Covaris). DNA fragments are ligated with dual-index adapters, and the amplified DNA libraries are size-selected using SPRIselect ® beads (Beckman Coulter Life Sciences, Indianapolis, IN, USA). ( 2 ) Shotgun libraries are enriched using a DNA probe based method of targeted capture enrichment for the whole mtgenome and eight categories of nuclear single nucleotide polymorphism SNP markers (426 SNPs). ( 3 ) The enriched samples are then sequenced on a NGS platform. ( 4 ) The nuclear SNP data is analyzed using NextGENe (SoftGenetics LLC, State College, PA, USA), while the mitochondrial data is analyzed using NextGENe, GeneMarker ® HTS (SoftGenetics LLC), and Mixemt [ 50 ].
Article Snippet: Nuclear DNA Sensitivity and Size Selection Study The single source male control DNA NA24129 (Coriell Institute for Medical Research, Camden, NJ, USA) was mechanically fragmented using the Covaris® M220 Focused-ultrasonicatorTM (Covaris, Woburn, MA, USA) to an average size of 175 bp with a range from 25 bp to 250 bp.
Techniques: Next-Generation Sequencing, Sequencing, Amplification