Library Validation for Next Generation Sequencing |
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Inaccurate quantification or poor normalization of next generation sequencing libraries prior to sequence amplification and detection can compromise results. Quantification and normalization are particularly important during multiplexing of samples or detecting intra-sample sequence variants.
How the PureGenome™ NGS Library Validator Kit Facilitates qPCR Analysis of Next Generation Sequencing Libraries:
- Three PureGenome™ NGS DNA Controls (150, 300, 600 bp) enable absolute molar quantification
- PureGenome™ Validator Primer Mix is optimized to ensure qPCR accuracy
- Highly specific for adapter-ligated libraries
- Sensitive and linear DNA detection
Benchmark Your Library Against Efficient Amplification of Standards:
Diluted DNA standards of known molar concentrations were tested with the PureGenome™ NGS library kit. High efficiency of qPCR amplification is represented independent of DNA size. Click image to enlarge.
Avoid Size-Bias Quantification!
Test Sample |
Competitor 1 Quantification (pM) |
Competitor 2 Quantification (pM) |
|---|---|---|
| 150bp DNA (1pM) | 1.06 | 0.06 |
| 300bp DNA (1pM) | 1.19 | 0.13 |
| 600bp DNA (1pM) | 0.06 | 0.34 |
Illumina sequencing ready fragments of 150, 300 and 600 bp were quantified using competitor kits following the manufacturer’s instructions. Competitor’s kits generate size-biased quantitation values. PureGenome™ NGS Validator Kit is uniquely optimized to minimize size bias using 3 PureGenome™ NGS DNA Controls and optimized PureGenome™ Validator Primer mix.