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VeriQuest® SYBR Green qPCR Master Mix (2X)

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Single tube master mix for Real-Time PCR, including SYBR Green, UDG, and ROX passive reference dye

Part # Description Unit Size
75600 40 RXN
VeriQuest SYBR Green qPCR Master Mix (2X)
1 ml for 40 reactions (50 µl)   
75600 200 RXN
VeriQuest SYBR Green qPCR Master Mix (2X)
5 ml for 200 reactions (50 µl)   
75600 400 RXN
VeriQuest SYBR Green qPCR Master Mix (2X)
2 x 5 ml for 400 reactions (50 µl)   
75600 1000 RXN
VeriQuest SYBR Green qPCR Master Mix (2X)
5 x 5 ml for 1,000 reactions (50 µl)   
75600 2000 RXN
VeriQuest SYBR Green qPCR Master Mix (2X)
10 x 5 ml for 2,000 reactions (50 µl)   

VeriQuest® SYBR Green qPCR Master Mix is supplied as a 2X pre-mixed formulation containing both ROX and UDG in an optimized buffer for quality results in real-time quantitative PCR assays. The proprietary reaction buffer and the hot start polymerase enhance SYBR Green-based qPCR amplification by reducing primer-dimer formation and non-template priming, increasing specificity and sensitivity. The master mix contains a chemically-modified Taq DNA Polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, Uracil-DNA Glycosylase (UDG or UNG), SYBR Green I, and ROX Passive Reference Dye in a proprietary reaction buffer.

Features:

  • One-tube master mix with ROX reference dye included
  • Same protocols and same primers for standard mode amplification
  • Consistency over a broad dynamic range: 7 orders of magnitude linear detection range with a correlation coefficient approximately equal to 1
  • Exceptional performance with challenging GC-rich regions
  • Detect less than 4 copies of target from genomic DNA and differentiate less than a two-fold difference
  • Contains dUTP and Uracil-DNA Glycosylase (UDG or UNG) for carryover contamination prevention
  • Hot start PCR for room temperature reaction setup
  • Stable at room temperature for 72 hours in a preassembled reaction

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USB VeriQuest SYBR Green qPCR Master Mix is supplied as a 2X pre-mixed formulation containing both ROX and UDG in an optimized buffer for quality results in real-time quantitative PCR assays. The proprietary reaction buffer and the hot start polymerase enhance SYBR Green-based qPCR amplification by reducing primer-dimer formation and non-template priming, increasing specificity and sensitivity.

 

  • Reproducible, consistent results while maintaining precision and efficiency
  • Enhanced amplification sensitivity and accuracy of low copy number detection
  • Exceptional performance with challenging templates
  • Ready to use Single Tube Replacement
  • Passive reference Dye and UDG included
  • Highly stable and easy to work with

 

Reproducible, consistent results while maintaining precision and efficiency
The convenience of a master mix formulation without sacrificing quality. VeriQuest SYBR Green qPCR Master Mix provides consistency over a broad dynamic template concentration range allowing 7 orders of magnitude linear detection. High precision in target quantification and discrimination of a 1.33 to 10-fold dilution series with VeriQuest SYBR Green qPCR Master Mix ensure accurate results.

Linear detection range of VeriQuest SYBR Green qPCR Master Mix. Real-time amplification plot from a 10-fold dilution series of a GAPDH synthetic target with starting amounts of 109 copies amplified in four replicate reactions using the ABI StepOneᆴ Real-Time PCR System.

 

Enhanced amplification sensitivity and accuracy of low copy number detection
Reliable detection from as little as 4 copies of a single copy gene and differentiation from less than a 2 fold difference.

High Precision in Target Quantification with VeriQuest SYBR qPCR Master Mix. Amplification plot from real-time PCR for a 1.33 to 10-fold dilution series of 10 ng to 1 ng of cDNAs reverse-transcribed from HeLa total RNA.

 

Exceptional performance with challenging templates
Even in high GC and AT rich regions VeriQuest SYBR Green qPCR Master Mix offers exceptional performance and high specificity so you have the confidence to verify your gene expression results.

Amplification of GC-rich target with VeriQuest SYBR Green qPCR Master Mix. Standard curves from a 10-fold dilution series of 100 ng to 10 pg human male genomic DNA amplified in four replicate reactions with VeriQuest SYBR Green qPCR Master Mix, BioRad iTaq Fast SYBR Green Supermix with ROX and ABI Power SYBR Green PCR Master Mix using the ABI 7500 PCR System for detection of a 71.2% GC-target amplicon (PROC, NT_022135.16)(3).

 

Melt curve from a 71.2% GC target amplicon. Human male genomic DNA was used.

 

Ready-to-use, Single Tube Replacement
The one tube master mix contains all necessary components: chemically modified Taq DNA polymerase, MgCl2, ultrapure nucleotides with an optimized dUTP:dTTP ratio, uracil-DNA glycosylase (UDG), SYBR Green I, and ROX as the passive reference dye in a proprietary reaction buffer. Simply add your DNA template, primers, and water and you can begin your qPCR reaction. For use in your new or existing protocols, on any leading PCR platform, providing easy transition from other master mixes.

 

Passive reference Dye and UDG included
ROX Passive Reference Dye is an inert dye, whose fluorescence does not change during the reaction. This signal normalization is necessary to correct for well-to-well differences that may occur due to artifacts such as pipetting errors or sample evaporation. Uracil DNA Glycosylase (UDG or UNG) and dUTP offer an option for carry-over contamination prevention from previous PCR amplifications. All are at optimized levels so there is no need to adjust concentrations.

 

Highly stable and easy to work with
VeriQuest SYBR Green qPCR Master Mix is stable at room temperature for 72 hours in a pre-assembled reaction and can be stored at 4°C for convenient handling, no lost time waiting for your master mix to thaw. Testing of 10 freeze-thaw cycles showed no loss in master mix performance. Ideal for high-throughput handling.

VeriQuest SYBR Green qPCR Master Mix Stability for Reliable Performance of High-Throughput Handling. Preassembled PCR reaction stability after 48 and 72 hour incubations at room temperature.

 

Inset defining Ct levels of 1ng & 10pg of cDNAs reverse transcribed from HeLa total RNA. GADPH was detected from preassembled PCR reactions incubated at room temperature for 48 and 72 hours.

 

Components:
VeriQuest SYBR Green qPCR Master Mix (2X):
40 reactions: 1ml
200 reactions: 5ml
400 reactions: 2x5ml
1,000 reactions: 5x5ml
2,000 reactions: 10x5ml
PCR Qualified Water 
Brief Protocol

 

References:

  1. VanGuilder, H. D., Vrana, K. E., and Freeman, W. M. (2008) Biotechniques 44 (5), 619-626.
  2. Longo, M. C., Berninger, M. S., and Hartley, J. L. (1990) Gene 93, 125128.
  3. Tewhey, R. et al. (2009) Nature Biotechnology 27, 10251031.
  4. Chittur, S. V. et al. (2009) A Follow-up on the Comparison of Different Priming Strategies for cDNA Synthesis by Reverse Transcriptase. Manuscript submitted for publication.

Applications:

  • Amplification from genomic DNA or cDNA input
  • Gene expression validation

  • Storage:

    Shipped on dry ice. -20°C for long-term storage. 4-8°C for short-term storage (≤3 months).

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Brochures

VeriQuest qPCR Solutions Brochure

PCR Tools Brochure

FAQs

USB VeriQuest qPCR MM FAQ

MSDS / Material Safety Data Sheets

VeriQuest SYBR Green qPCR Master Mix (2X) SDS

Product Spotlight

VeriQuest SYBR Green qPCR Master Mix Product Spotlight

Protocol, Brief

VeriQuest SYBR Green qPCR Master Mix (2X) Brief Protocol

Tech Tips

Instrument Compatibility Guide Tech Tip

PCR product selection guide

Tutorials

Primer Selection Guidelines Utilizing VeriQuest qPCR for Microarray Validation Technical Note

White Papers

Cytoscan Copy Number Confirmation with SYBR Green qPCR White Paper

Concordance of HTA Array and Real Time qPCR Results White Paper

Concordance of Microarray and RT-PCR Results White Paper

Other

EB 2014 Concordance of HTA Array Poster

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