EasySeq™ Human DNA Sample Identification

Complex sample preparation workflows and challenges handling multiple Next-Generation Sequencing (NGS) samples make misidentification of samples at any stage of the analytical process a recognizable concern. A method for independent confirmation of sample identity is therefore highly desirable, preferably using an identification method that is intrinsic to the WGS/WES data of the biological sample and simple to incorporate into the existing NGS workflow (Figure 1).

EasySeq™ Human DNA Sample Identification Kits, powered by patented Reverse Complement PCR (RC-PCR) technology, facilitate an effective end-to-end identification and tracking of independent samples in a single targeted sequencing assay that is purpose-designed for fast and efficient lab processing and, moreover, high assay sensitivity, specificity, safety and discriminatory power required in human genetics analysis. Its one-tube single reaction RC-PCR workflow greatly reduces the amount of hands-on steps and the associated risks of pipetting errors, sample swaps and cross-contamination, safeguarding sample identity and data integrity.

EasySeq™ Human DNA Sample Identification Kits are complementary to both Whole Exome and Whole Genome Sequencing Sample workflows. Sample IDs are matched with WES/WGS data and, as a result, data integrity and validity can be confirmed.

Another application of the EasySeq™ Human DNA Sample Identification Kit is the conformation of cell line authentication in Human Cell-line Culturing.

EasySeq™ Human Sample Identification Kits offer laboratories the simplest and safest workflow for NGS sample identification available in the market. RC-PCR in combination with the optimized panel provides a number of distinctive features and benefits that help safeguarding sample identity and data integrity:

Confidence in test results

• MAF SNPs selected in combination with RC-PCR kinetics provide high discrimination power

• UDIs of 10 bp increase discriminatory power and help to prevent index hopping

• Minimizing pipetting error and preventing sample swaps: correct data interpretation and the right critical samples are matched

• Sample tracking dye in pre-spotted Unique Dual Indexing plates ensures accuracy

• Also validated with challenging cfDNA and FFPE DNA samples

• Unique RC-PCR kinetics: high-target specificity and coverage uniformity (optimized read depth balance) from low DNA input

• Blood Direct Kit: include the DNA isolation step in the validation process, to exclude possible sample swabs or contamination

Cost-efficient workflow

• Breakable universal index (IDX) plates ensure optimal usage, minimizing waste

• One closed-tube, single reaction workflow with simultaneous indexing and target amplification reduces labor time

• Single tube sample pooling for library clean-up significantly reduces usage of magnetic beads and consumables

• Well-balanced read distributions maximize sequencing instrument flow cell capacity

• Costly sample re-runs are prevented by capturing misidentified samples

Choice and Flexibility

• Complementary to both Whole Exome and Whole Genome Sequencing workflows

• Compatible with various Illumina® platforms

• A variety of 96-well breakable UDI plates available for matching your sample workload, facilitating up to 768 samples per run

EasySeq™ Human DNA Sample Identification Kits are the improved version of the widely adopted EasySeq™ Human WES WGS Sample Tracking Kit (RC-HEST096), comprising an optimized panel of 40 exonic targets, the genotypic profile which can be utilized to extract intrinsic identifiers from the human exome and genome: 37 exonic single  nucleotide polymorphisms (SNPs) with high minor allele frequency (MAF), and Amelogenin X - Y and TXLNGY for gender determination (Table 1).

The NimaGen EasySeq™ Human DNA Sample Identification Kits offer a simple, seamlessly integrated NGS approach to ensure sample identification and tracking from an early step in the workflow, using either extracted DNA or running multiplex RC-PCR directly on the blood sample (fresh or frozen blood, dried bloodspots). NimaGen offers two Illumina® compatible versions of the EasySeq™ Human DNA Sample Identification Kit:

• EasySeq™ Human DNA Sample Identification Kit supports an NGS driven human DNA sample tracking and identification strategy for human genetics, using a single-panel multiplex RC-PCR reaction, based on the analysis of DNA isolates.
• EasySeq™ Human DNA Sample Identification - Blood Direct supports an NGS driven human DNA sample tracking and identification strategy for human genetics, using a single-panel multiplex RC-PCR reaction directly from blood.