QIAGEN Website    Quick Order    Online Seminar    Contact    My Account
Home  >  Products and Services  >  GeneRead NGS   >  GeneRead Library Quant Kit

GeneRead Library Quant Kit

NGS Platform

Passive Reference Dye

Catalog #

Price and Ordering

Illumina

None

NGTF-ILZ-N

Fluorescein

NGTF-ILZ-F

Rox

NGTF-ILZ-R

FAST with ROX (for RGQ)

NGTF-ILZ-Q

Ion Torrent

None

NGTF-ITZ-N

Fluorescein

NGTF-ITZ-F

Rox

NGTF-ITZ-R

FAST with ROX (for RGQ)

NGTF-ITZ-Q

Each kit is enough for 500 reactions and includes necessary PCR Mastermix

One of the most important factors in a next-generation sequencing experiment is accurate quantification of the prepared library. The GeneRead DNAseq Library Quant Array uses real-time PCR to quantify NGS libraries by specifically quantifying DNA molecules with adaptors at both ends, which are the only amplifiable molecules during emulsion PCR (Ion Torrent platform) or bridge PCR (Illumina platform), and therefore provides highly accurate quantification of amplifiable library molecules. The high sensitivity of real-time PCR allows quantification of libraries with very low concentrations, even below the detection threshold of conventional spectrophotometric methods.

The GeneRead Library Quant Kit provides NGS platform specific DNA standard and PCR primer assay for reliable and convenient quantification of NGS library. An optimized GeneRead qPCR SYBR Green Mastermix is included in the kit to provide superior performance.

Why GeneRead Library Quant Kit?

  • Simple protocol, high sensitivity and wide dynamic range
  • Data analysis Excel sheet automatically calculates library concentration
  • Compatible with all major NGS platforms and qPCR instruments

How it works


Figure 1.† GeneRead DNAseq Library Quant Array workflow.


Performance Consistency


Figure 2. Reliable NGS library quantification with minimal variability of DNA standards from lot-to-lot.
The DNA standard for Illumina MiSeq platform from three different lots were used to prepare five sequential 10-fold dilutions. The diluted DNA standards were mixed with library specific PCR primer assay and GeneRead qPCR SYBR Green mastermix (from three different lots), and subjected to real-time PCR. The minimal variation in Ct values for diluted DNA standards from three different lots confirms the reliability of GeneRead Library Quant System.

Application Data

GeneRead Library Quantification System enables quantification of libraries with concentration below detection limit of conventional methods


NGS libraries were constructed for Ion Torrent PGM platform using Human Comprehensive Cancer GeneRead DNAseq Gene Panel from universal genomic DNA (NGS-L1) and genomic DNA isolated from FFPE lung carcinoma sample (NGS-L2) and subjected to quantification by High Sensitive DNA Assay Kit (BioAnalyzer). BioAnalyzer can quantify only NGS-L1 whereas concentration of NGS-L2 was too low to quantify (A). GeneRead Library Quant Systemís high sensitivity and broad dynamic range enables the quantification of both NGS-L1 and NGS-L2. The result of library quantification for NGS-L1(blue arrow) and NGS-L2(red arrow) with 1:2000 dilution is shown in figure B. Amount of NGS-L1 and NGS-L2 amount obtained by both methods are summarized in Figure C.

NGS System Home
DNAseq Targeted Panels
DNAseq Mix-n-Match Panels
DNAseq Custom Panels
DNAseq Panel Mastermix
GeneRead Library Quantification
Library Quantification Mastermix