Molecular Biology - Next Generation Sequencing - Library Validation

Next Generation Sequencing Library Quantification and Validation

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.

The PureGenome^™ Next Generation Sequencing Library Validator Kit enables highly accurate qPCR quantification of Illumina platform-compatible NGS libraries. The kit includes three DNA standards covering optimal library size ranges and a primer mix exhibiting validated amplification efficiency.

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

Order PureGenome NGS Library Validator Kit)

Flexible, open platform allows use with any qPCR protocol using SYBR^® Green technology.

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.