Massively parallel sequencing for the forensic scientist – sequencing archived amplified products of AmpFlSTR Identifiler and PowerPlex Y multiplex kits to capture additional information

Abstract

In the last few years the cost and ease of massively parallel sequencing (MPS) has reduced dramatically to the point that it can now be considered as a tool for use in forensic case work. An important consideration for the implementation of any new forensic technology is the ability to remain compatible with previous technology. During this study we sequenced the amplicons of two commercial forensic short tandem repeat (STR) multiplexes AmpFlSTR Identifiler and PowerPlex Y using the Illumina MiSeq and Ion PGM Sequencer (Life Technologies) and characterised the sequence data from a forensic perspective. Using the MPS data from both platforms we determined the STR genotypes of forensic samples and found previously undocumented sequence variation in seven STR alleles. By characterising features of the DNA sequence profiles, such as stutter and locus imbalance we identified areas for future development that will be needed prior to casework implementation. The rapid development of this technology has meant many in the forensic community have been ‘left behind’. We also provide an explanation, for forensic scientists, of what is happening at the different stages of the MPS workflow, from library preparation through to bioinformatics, and how this may affect the results.

Keywords:

• massively parallel sequencing
• short tandem repeats
• Ion PGM Sequencer
• Illumina MiSeq
• forensic science

Additional information

Funding

This work was supported by Institute of Environmental Science and Research.