Absolute quantification, using sealed-chip technology, for a reliable method and precise, sensitive data.
Next-generation sequencing (NGS) libraries can be quantified with minimal sample handling and without the need to generate a standard curve using digital PCR. This method enables accurate and precise library quantification, a critical step in both the Ion Torrent™ and Illumina® workflows, allowing for maximizing sequencing yields downstream. To achieve this high degree of precision, a TaqMan® Assay, designed to span both the forward and reverse adapters specific to each library, is available.
This approach limits quantification to library constructs that contain both adapter sequences. Ultimately, using digital PCR to quantify NGS libraries decreases overall sequencing costs by ensuring an accurate quantification upfront, minimizing the need to re-run or repeat sequencing of samples.
|Sample name||Library type (size)||Concentration determined by digital PCR||Templated beads pre-enrichment|
|L7734||Fragment (276 bp)||3.32 nM||12.1%|
Fragment (489 bp)
|L2444||Fragment (322 bp)||11.03 nM||10%|
Table 1: Correlation of QuantStudio™ 3D Digital PCR data with that of the Ion OneTouch™ 2 data. These quality-control data were generated for four Ion Torrent™ libraries using flow cytometry prior to enrichment.
Peter Schweitzer, PhD, Director of the Genomics Facility at Cornell University, Ithaca, New York
Application Note: Precise quantification of Illumina® libraries on the QuantStudio® 3D Digital PCR System Application Note: Precise Quantification of Ion Torrent™ Libraries on the QuantStudio™ 3D Digital PCR System Demonstrated protocols in Digital PCR Community
Product Bulletin: QuantStudio™ 3D Digital PCR System
User Guide: QuantStudio™ 3D Digital PCR System
Digital PCR Experiment Design Guide
Poster: Precise Quantification of Next Generation Sequencing Ion Torrent™ Libraries with the QuantStudio™ 3D Digital PCR Platform
For Research Use Only. Not for use in diagnostic procedures.