Next Generation Sequencing in Forensic Science: A Primer addresses Next Generation Sequencing (NGS) specific to its application to forensic science. The book opens with a history of human identity approaches including VNTR, RFLP, STR, and SNP DNA typing.
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Next Generation Sequencing in Forensic Science: A Primer addresses Next Generation Sequencing (NGS) specific to its application to forensic science.
The book opens with a history of human identity approaches including VNTR, RFLP, STR, and SNP DNA typing. It next discusses the history of sequencing for human DNA typing including: Sanger sequencing, SNaPshot, pyrosequencing, and principles of next generation sequencing. Readers are expected to have as basic understanding of molecular and cellular biology and DNA typing. Chapters present an overview of the forensically-focused AmpliSeq, ForenSeq, Precision ID, PowerSeq and QIAseq panels for human DNA typing using autosomal, Y and X chromosome STRs and SNPs run on the MiSeq FGx and Ion Torrent System. Using these, that authors outline the steps included in DNA extraction and DNA quantitation that are performed prior to preparing libraries with the NGS kits.
The second half of the book details the implementation of ForenSeq and Precision ID to amplify and tag targets to create the library, enrich targets to attach indexes and adaptors, perform library purification and normalization, pool the libraries, and load samples to the cartridge to perform the sequencing on the instrument. Coverage addresses the operation of the MiSeq FGx and Ion Chef including creating a sample list, wash steps, performing NGS, understanding the run feedback files from the instrument, and troubleshooting. ForenSeq and Precision ID panel data analysis are explained including how to analyze and interpret NGS data and output graphs and charts. The book concludes with the mitochondrial chromosome and mitochondrial DNA (mtDNA) sequencing and SNPs analysis including the issue of heteroplasmy. The final chapters review applications of microbial DNA for forensics, NGS in body fluid analysis, and challenges and considerations for future applications.
Next Generation Sequencing in Forensic Science is the first book to prepare practitioners to utilize and implement this new technology into their lab for casework, highlighting early applications of how NGS results have been used in court. The book can also be utilized for upper-level undergraduate and graduate students taking courses focused on NGS concepts.