Description
ForenSeq Kintelligence HT is the first targeted sequencing workflow designed for unidentified persons and remains or large-scale kinship projects. Targeting 10,230 SNP markers specifically curated for kinship determination, the assay is optimized to work with challenging samples such as bones. Leveraging the unique dual index (UDI) plate allows the library preparation workflow processing of up to 96 postmortem (PM) or antemortem (AM) samples before moving on to sequencing. The ForenSeq Kintelligence HT workflow enables a higher level of multiplexing that balances throughput with the closer kinship determinations typically required for unidentified persons and remains. Detect at least third-degree kinship with 100% sensitivity and specificity when sequencing up to 12 postmortem samples or 36 antemortem samples per MiSeq FGx run. Analyze data in the Universal Analysis Software (UAS) using the preconfigured ForenSeq Kintelligence HT Analysis Method. Utilize the new UAS database exclusively for Kintelligence HT featuring local database storage, kinship comparison of postmortem to antemortem samples, and likelihood ratios for relationship comparisons.
The ForenSeq Kintelligence HT kit is the first forensic assay to combine high throughput targeted SNP sequencing and local kinship analysis database for the large-scale identification of human remains. The fully integrated workflow includes optimized chemistry for highly degraded and contaminated samples, higher level multiplexing that balances throughput with closer kinship determinations, and a fit-for-purpose analysis software and kinship database that can be used on a local database to maintain the privacy of the families concerned. ForenSeq Kintelliegnce HT has optimized workflows for both postmortem (PM) and antemortem samples (AM) with sequencing capabilities up to 12 PM libraries and 36 AM libraries obtaining at least third degree relationship. High throughput, fit for purpose workflow: Kinship power beyond STRs Reliably determine relationships out to the third degree, even with degraded samples. Sensitive solution for low input, low quality samples Sequence compromised samples on a forensically validated system for easily accessible results. A local database for kinship analysis Pair two familiar workflows, ForenSeq and GEDmatch PRO algorithms for kinship testing Feature Details Kit configuration 96 reactions Number of SNPs 10,230 Mean amplicon size < 150 bp Total library prep time 10 hours Hands-on library prep time < 3 hours Sequencing time 28 hours Sample type tested Blood, bone, buccal swabs, rooted hair, teeth, and semen Antemortem samples Postmortem samples Recommended input 1 ng 500pg Multiplexing capacity 36 libraries per run 12 libraries per run
A robust PCR and primer design enables a single SNP multiplex, maximising kinship power. The small average amplicon size of < 150 bp improves amplification efficiency and facilitates recovery and analysis of degraded DNA. Additionally, the ForenSeq Kintelligence HT Kit includes 96 unique dual index (UDI) adapters, which attach unique identifying sequences to each end of the library for optimum data recovery. ForenSeq Kintelligence HT queries 10,230 SNPs carefully selected to maximise kinship power. Category Number of SNPs Percentage of total Ancestry SNPs 56 0,5% Identity SNPs 94 1% Kinship SNPs 9867 96% Phenotype SNPs* 22 0,2% X-SNPs 106 1,2% Y-SNPs 85 0,9% *Two SNPs overlap the ancestry and phenotype categories and are counted in the phenotype category only. The ForenSeq Kintelligence HT Kit utilizes the same SNPs found in the original ForenSeq Kintelligence Kit, including the 9867 kinship SNPS, as well as 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. 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 HT excludes the SNPs with known medical associations or low minor allele frequencies to limit privacy concerns and protect genetic health data. 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 HT inputs are compatible with DNA extraction methods common in forensic laboratories. Library preparation leverages the same chemistry backbone that is foundational to the ForenSeq library prep portfolio. ForenSeq Kintelligence HT reagents enable preparation of up to 96 dual-indexed, human-specific libraries in 10 hours, with < 3 hours of hands-on time. Pooling libraries recommendation for sequencing are 12 PM samples or 36 AM samples. Sequencing is completed in 28 hours and analysis in 1 hour, including upload to the local UAS database for kinship and likelihood analysis. The fully integrated NGS workflow delivers results. In addition to providing streamlined 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. The 96-reaction kit includes master mixes for amplification, purification beads for cleanup and is supplied with straightforward protocols, featuring six safe stopping points.
