Digital PCR Library Quantification Kits

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Overview

QX200™ Droplet Digital™ Brochure

Droplet Digital PCR (ddPCR™) is Bio-Rad's unique digital PCR technology.

  • Unrivaled precision provides absolute quantification of target DNA or RNA molecules without the use of standard curves
  • Addresses the lack of scalable and practical technologies for digital PCR implementation

The QX200 ddPCR system makes this powerful technology accessible to you for unveiling previously unattainable
research levels.

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QX200™ Droplet Digital™ (ddPCR™) Data
Detection of BRAF V600E Mutation

High sensitivity of the QX200 system allows quantification of BRAF V600E mutation in the presence of wild-type DNA using PrimePCR ddPCR assays. A, 2-D fluorescence amplitude plot generated by QuantaSoft™ software shows triplicate wells of a mixed mutant:wild-type sample. The black cluster on the plot represents the negative droplets, the green cluster represents the droplets that are positive for wild-type DNA only, the blue cluster represents the droplets that are positive for mutant DNA only, and the orange cluster represents the droplets that are positive for both mutant and wild-type DNA. B, 2-D fluorescence amplitude plot shows three replicates of a wild-type-only sample. C, fractional abundance plot shows the percentage frequency (orange markers) of the mutant DNA in a wild-type DNA background. The blue markers indicate the concentration of mutant DNA (copies/µl) and the green markers indicate the concentration of wild-type DNA (copies/µl) in each of three replicate samples. All error bars generated by QuantaSoft software represent the 95% confidence interval.



Detection of Herpes Simplex Virus and β2 Microglobulin

ddPCR enables precise and reproducible detection of herpes simplex virus 1 (HSV-1) and ß2 microglobulin (B2M) targets in a duplex assay. A, 2-D fluorescence amplitude plot shows three merged replicate samples of 10,000 copies/well of HSV-1 duplexed with 20,000 copies/well of Β2M. The black cluster on the plot represents the negative droplets, the green cluster represents the droplets that are positive for HSV-1 only, the blue cluster represents the droplets that are positive for B2M only, and the orange cluster represents the droplets that are positive for both HSV-1 and B2M targets. B, concentration plot shows merged triplicate wells across an HSV-1 sample dilution series from 0 to 40,000 copies in a constant background of 20,000 copies (66 ng human genomic DNA) of B2M. The blue markers indicate HSV-1 copies/µl and the green markers indicate B2M copies/µl. All error bars generated by QuantaSoft software represent the 95% confidence interval.



Detection of Copy Number for
Myelocytomatosis Oncogene and β-Globin

The QX200 system provides superior resolution with copy number (CN) calls from CN1 to CN13. A, 2-D fluorescence amplitude plot shows duplicate wells of a CN6 sample duplexed with myelocytomatosis oncogene (MYC) and β-globin (HBB) assays. The black cluster on the plot represents the negative droplets, the green cluster represents the droplets that are positive for MYC only, the blue cluster represents the droplets that are positive for HBB only, and the orange cluster represents the droplets that are positive for both MYC and HBB targets. B, copy number plot shows samples ranging from CN1 to CN13 with precise duplicate well values. All error bars generated by QuantaSoft software represent the 95% confidence interval. NTC, no template control.



Validation for Next-Generation Sequencing

ddPCR provides nearly 1,000-fold greater sensitivity and an amplification bias–free approach compared to RNA-Seq for validation. A, concentration plot shows a twofold dilution of cDNA generated using Bio-Rad’s iScript™ advanced cDNA synthesis kit for RT-qPCR. Two independent measurement sets were made: one at a high concentration range and one at a lower range, with four points overlapping. B, table shows comparative results between ddPCR and MiSeq. ddPCR detected thousands of copies per well with low-abundance transcripts (TBP and GUSB genes) using an input of 100 ng total RNA, which demonstrates enhanced sensitivity. The MiSeq sequencer detected only single-digit reads per kilobase per million reads (RPKM) for these transcripts. ddPCR, Droplet Digital PCR.



Gene Expression Analysis of Stem Cell Markers

High sensitivity and precision of the QX200 system allow reliable detection of small fold changes with rare transcripts. The TGFβ signaling pathway plays a role in ovarian stem cell regulation, and the addition of TGFβ will increase the number of stem cells in the population, increase the expression of putative stem cell markers, and decrease terminal differentiation markers like E-cadherin (ECAD). Data show expression levels of cKIT, CD133, OCT3/OCT4, and ECAD genes across ten samples with and without TGFβ treatment, normalized to PPIA in this study. Reproducible detection of cDNA copies as low as 0.5–1 target/µl in a 20 µl reaction was achieved. S, sample.


PrimePCR™ ddPCR™ Assays Brochure

PrimePCR™ Assays for
Droplet Digital™ PCR

Expertly designed and validated assays

PrimePCR™ ddPCR™ Assays are validated assays that enable precise quantification without a standard curve.

  • Generate consistent and reproducible results
  • Utilize universal cycling conditions and primer/probe strategy
  • Guarantee performance

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Precise quantification of next-generation sequencing (NGS) libraries is critical for the efficient use of NGS platforms. Bio-Rad's digital PCR platform and library quantification kits can be easily incorporated into the NGS library preparation workflow to precisely quantify and balance sequencing libraries on the Illumina or Ion Torrent Platforms.

The ddPCR library quantification assay generates data plots that are rich with qualitative library information, a feature not available with other current methodologies.

When paired with the QX200™ Droplet Digital™ PCR System, these kits allow accurate quantification and provide a qualitative measure of the DNA library prior to sequencing on Illumina (186-3040) and Ion Torrent (186-3041) Platforms.

Features of Digital PCR Library Quantification Kits

  • Contain all the necessary components to create droplets and quantify NGS libraries
  • Provide information about library quality, such as adapter dimers, and indicate library insert size
  • Provide more efficient and consistent loading of libraries for sequencing runs
  • Enable balancing of pooled library samples
  • Optimize the use of consumables, labor, and instrument time for the highest productivity with NGS

Related Products

Bio-Rad offers additional digital PCR supermixes including:

Additional Equipment

These digital PCR assays require the use of the following instruments and accessories:

Storage at –20°C Guaranteed up to 12 months in a constant temperature freezer (multiple freeze thaws not recommended)
ddPCR™ Library Quantification Kit for Illumina TruSeq

186-3040
200 x 20 µl reactions, includes droplet PCR supermix and ddPCR library quantification assay, for quantification of Illumina TruSeq libraries using the QX200™/QX100™ ddPCR™ systems

List Price:   $511.00
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ddPCR Library Quantification Kit for Ion Torrent
ddPCR™ Library Quantification Kit for Ion Torrent

186-3041
200 x 20 µl reactions, includes ddPCR supermix for probes (no dUTP) and ddPCR library quantification assay, for quantification of Ion Torrent AmpliSeq and RNA libraries using the QX200™/QX100™ ddPCR™ systems

List Price:   $499.00
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Number Description Options
6402 ddPCR Library Quantification Kit for Illumina TruSeq Flier, Rev B Click to download
6490 ddPCR Library Quantification Kit for Ion Torrent Flier, Rev A Click to download
10031986 ddPCR Library Quantification Kit for Illumina TruSeq Product Insert, Rev C Click to download
10033630 ddPCR Library Quantification Kit for Ion Torrent Product Insert, Rev B Click to download
6450 Droplet Digital PCR Publications List, Rev E Click to download
6423 Droplet Digital PCR: Ultra-Sensitive Validation Technology for RNA-Seq Application Note, Rev A Click to download [ Add to Cart (Free) ]
10023997 Instruction Manual, PX1 PCR Plate Sealer, Rev A Click to download
6495 Bio-Rad’s ddPCR™ Library Quantification Kit for Ion Torrent Enables Accurate Library Quantification and Excellent Balancing of Pooled Libraries, Rev A Click to download