ForenSeq® DNA Signature PLUS Kit

Detailed description

Operating principle

Based on amplicon technology, ForenSeq® DNA Signature Plus Kit generates dual-indexed DNA libraries for sequencing on the MiSeq FGx platform. Each sample is combined with a mixture of specific primers labelled for each target sequence. A first multiplex PCR amplifies the selected targets. A second PCR incorporates unique dual indexes (UDI) for each sample and enriches the amplified products. The resulting libraries are purified and normalised using magnetic beads and are finally combined into a pool ready for sequencing. The kit includes all the reagents required for every stage of library preparation, as well as NA24385 control DNA.

This kit enables STR and SNP markers to be amplified in the same reaction, generating a profile that combines different categories of forensic markers and providing two content options (DPMA or DPMB) depending on the objective of the study. The DPMA mix amplifies 27 autosomal STRs, 24 Y-STRs, 7 X-STRs and 94 identity-informative SNPs; the DPMB mix additionally provides 22 phenotype-informative SNPs (eye colour and hair colour) and 56 biogeographical ancestry-informative SNPs.

ForenSeq® DNA Signature Plus Kit is suitable for different types of starting material. It has been validated for purified DNA samples, buccal swab lysates and direct punches from FTA cards. An optional PCR buffer, ForenSeq® Enhanced PCR1 Buffer (ePCR1), is also available and is specifically designed for samples containing inhibitors such as humic acid or haematin.

The results obtained are analysed using a specific UAS module included on a dedicated server. The server running the independently installed UAS software receives the data and performs the analysis corresponding to the assay: FASTQ generation, alignment, variant calling and reporting of results.

The MiSeq FGx instrument operates with two reagent cartridges of different capacities, enabling sequencing runs to be optimised according to the number and nature of the samples. Recommendations for forensic cases take into account potential mixtures, less than 1 ng of available DNA or partial degradation. For reference samples, the multiplexing capacity is higher.

Applications

The high number of markers, their different nature and their distribution across two different primer mixes make ForenSeq® DNA Signature Plus a highly versatile kit that can support forensic geneticists in different applications:

Intended users

Forensic genetics, human identification and kinship testing laboratories requiring a high number of markers to resolve complex identification or kinship cases.

Key aspects