RC-PCR powered EasySeq™ NGS Library Prep for 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.
Confidence in Data Integrity and Validity
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
• 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
Intrinsic Identifiers from the Human Exome and Genome
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.