Verogen ForenSeq Kintelligence Kit

Target 10,230 SNP markers explicitly curated for extended kinship outcomes with the ForenSeq Kintelligence Kit. The only sequence-based assay designed for forensic genetic genealogy (FGG), ForenSeq Kintelligence minimizes privacy concerns by excluding medically informative and minor allele frequency SNPs found in whole-genome sequencing (WGS) or array-based workflows. Robust PCR and primer designs allow for a single SNP multiplex that works across global populations.

ForenSeq Kintelligence is an integrated solution for forensic genetic genealogy (FGG). Libraries are prepared, sequenced and analyzed in a single, streamlined workflow for efficient assessment of forensically relevant SNPs. Developed according to existing STR-based methods, with input from on-market methods and established research and databases, the kit provides laboratories targeted sequencing and accurate analysis from forensic DNA samples, including those that are low-input and highly degraded. By applying a curated SNP set, a low input requirement, and a forensically validated workflow while building on established methodology, ForenSeq Kintelligence offers the opportunity to advance beyond dead ends and find resolution for cold cases, missing persons and unidentified human remains.

• Sequence compromised samples on a validated system for easy access to results
• Curated content design ensures data privacy with a SNP set selected for forensic relevance
• Keep evidence and associated data in your laboratory and retain full control
• Fully integrated and supported to pair two familiar workflows, ForenSeq and GEDmatch

Cold case samples and missing persons samples are often degraded due to advanced age, exposure to environmental elements and contaminants, and variable extraction and storage procedures. Accordingly, Verogen designed the ForenSeq Kintelligence Kit to deliver high recovery rates from compromised samples, performing studies analyzing degraded and inhibited casework-type samples and conducting a sensitivity assessment with control samples so that the kit reproducibly generates SNP calls across a range of input DNA amounts.

ForenSeq Kintelligence delivered high performance in mock casework studies that generated high call rates from degraded blood samples, teeth samples inhibited by calcium and contemporary bone samples subjected to the insults of cremation, burning and embalming. The sensitivity assessment evaluated control DNA at inputs ranging from 5 ng to 50 pg. The inputs ranging from 5 ng to 250 pg all achieved a call rate of 100%. Performance was similarly high at 100 pg and 50, at 99.9% and 99.6%, respectively (see figure “A sensitivity titration of control DNA across a range of inputs”). High average coverage of approximately 1500× across target sites allows these high call rates and ensures confident calls.

Given exposure to natural elements and prolonged degradation through age, cold case samples are prone to failing quality control checks and yielding poor coverage. This results in low DNA recovery and impacts allele call rates. To maximize recovery, ForenSeq Kintelligence buffers tolerate many common inhibitors including hematin, humic acid, indigo and tannic acid, and microbial contaminants such as E. coli. For 1 ng gDNA samples spiked with 10 ng microbial DNA, Verogen observed a SNP recovery rate of 100%. Control samples inhibited with spike-ins of significant amounts of other common forensic inhibitors demonstrated a high average call rate of 99.2%, comparable with control samples (see figure “An evaluation of inhibitor performance“). The high average call rate enables deep coverage of samples regardless of low input.

A robust polymerase chain reaction (PCR) and primer design enables a single SNP multiplex, eliminating the need for multiple tests. The small average amplicon size of < 150 bp improves amplification efficiency and facilitates recovery and analysis of degraded DNA. Additionally, the ForenSeq Kintelligence Kit includes six unique dual index (UDI) adapters, which attach unique identifying sequences to each end of the sample for optimum data recovery.

ForenSeq Kintelligence queries 10,230 SNPs that have been carefully selected to support targeted sequencing for FGG.

These SNPs overlap with relevant markers in the Illumina Infinium CytoSNP-850K BeadChip and Infinium Global Screening Array and are cross‑referenced against the Genome Aggregation Database (gnomAD) v3.0 and Single Nucleotide Polymorphism database (dbSNP) v151 for robust performance across global populations. Importantly, ForenSeq Kintelligence excludes the SNPs with known medical associations or low minor allele frequencies to limit privacy concerns and protect genetic health data.

The design of ForenSeq Kintelligence was influenced by ForenSeq DNA Signature Prep to include all biogeographical ancestry, identity and phenotype SNPs validated for investigative lead generation. Also included are informative Y-SNPs and X-SNPs to help with lineage and biological sex determination, respectively.

An optimized buffer system tolerates a range of inhibitors commonly found in forensic samples, such as calcium, indigo and humic acid, generating a > 98% call rate. Assay design provides a high level of process integrity and data quality so you can trust the analysis of even the most challenging forensic samples.

ForenSeq Kintelligence inputs are compatible with DNA extraction methods common in forensic laboratories. Library prep leverages the ForenSeq chemistry backbone that is foundational to the Verogen library prep portfolio. ForenSeq Kintelligence reagents enable preparation of up to 12 dual-indexed, human-specific libraries in 8 hours with only 1 hour and 55 minutes of hands-on time. Sequencing is completed in 28 hours and analysis in 1 hour for a fully integrated NGS workflow that delivers results, including a GEDmatch PRO report that users can directly upload to the database, in fewer than 40 hours. If a match exists, GEDmatch PRO produces results in fewer than 2 days.

In addition to providing rapid library prep that includes normalization, the ForenSeq chemistry offers a high degree of flexibility for sample sources such as buccal swabs, blood, bones and teeth, while an input volume of 25 μl enhances support for degraded and inhibited samples. The 12-reaction kit includes master mixes for amplification, purification beads for cleanup and straightforward protocols featuring six safe stopping points.

FGG combines microarray genotyping or whole‑genome sequencing (WGS) methods of DNA profile generation with comparisons of genetic relatives in genealogy databases such as GEDmatch, the largest database of voluntarily submitted DNA profiles for forensic comparisons. Comparisons made in a database allow genealogists to construct a family tree using census records, vital records, obituaries and newspaper archives, then trace the source of the DNA through the family tree.

Microarray and WGS-based methods cannot support the full spectrum of biological samples associated with forensic cases. These samples are often degraded or inhibited due to source material age or environmental exposure, resulting in low‑quality data that limits FGG. Microarrays and WGS also produce large amounts of medically relevant single nucleotide polymorphism (SNP) data that are not applicable to forensic applications, raising concerns about genetic data privacy and usage. The ForenSeq Kintelligence Kit is the only FGG assay fit for forensic purpose. Designed for sequencing on the National DNA Index System (NDIS)‑approved MiSeq FGx Sequencing System, data analysis in Universal Analysis Software (UAS), together with long‑range kinship analysis in GEDmatch PRO, ForenSeq Kintelligence and a Verogen STR assay can take you from DNA to a confirmed identity.

The end-to-end workflow of the ForenSeq Kintelligence Kit is based on the established ForenSeq process, enabling a diverse range of DNA analysis on a single, proven platform. The ForenSeq Kintelligence Kit links seamlessly with the ForenSeq Kintelligence Analysis Module in UAS for rapid data assessment and single-click report generation.

We offer superior support across the entire workflow, from library prep to sequencing to analysis. Our experienced team provides comprehensive service coverage for your equipment and software, validation plans, and implementation guidance so you can quickly operationalize your workflows with ease.