qPCRBIO SyGreen Mix Hi-ROX – Witec AG

Art No | PB20.12
from 53.00 CHF

qPCRBIO SyGreen Mix is designed for fast, highly sensitive and reproducible real-time PCR.


The antibody-mediated hot start system prevents the formation of primer dimers and non-specific products leading to greatly improved reaction sensitivity and specificity.

qPCRBIO SyGreen Mix combines a proprietary non-inhibiting intercalating dye (detected in the FAM channel) with the latest advances in polymerase technology and buffer chemistry to give you fast, highly sensitive and reproducible real-time PCR.

qPCRBIO SyGreen Mix can be used to reliably quantify any DNA template including genomic, cDNA and viral sequences, and is able to detect extremely low copy number targets with the highest efficiency.

  • Rapid extension rate for early Ct values
  • Market-leading sensitivity – increased limit of detection
  • Specific amplification from complex templates including GC-rich sequences
  • Non-PCR inhibiting intercalating dye
  • Also available as an easy-to-see blue mix
  • Compatible with all standard and fast cycling real-time instruments

The 2x mix uses a proprietary intercalating dye that does not inhibit PCR, unlike other popular fluorescent dyes. The same developments that improve the sensitivity and consistency of qPCRBIO SyGreen Mix in standard cycling conditions also allow for industry-leading performance in fast and ultra fast cycling conditions.


  • Absolute quantification
  • Relative gene expression analysis
  • High-throughput qPCR from genomic, cDNA and viral sequences
  • Detection of extremely low copy number targets
  • Crude sample PCR

Use our qPCR Selection Tool to find out which ROX variant is compatible with your instrument.


The kit should be stored between -30°C and -15°C. Avoid prolonged exposure to light. If stored correctly the kit will retain full activity for 12 months. The kit can be stored at 4°C for 1 month. The kit can go through 30 freeze/thaw cycles with no loss of activity.

Amplification of Beta-2 Microglobulin using qPCRBIO SyGreen Mix (purple curves).

Amplification curves are shown in the top panel of set a and set b, melt curves are shown in the middle panel and the efficiencies of amplification are shown in the bottom panel.

A direct, on-plate comparison was performed with the competitors identified in the top panel of each set (grey curves). 5 serial dilutions of mouse cDNA template were used in a total reaction volume of 10µL. Cycling conditions were those recommended by each of the competitors.

qPCRBIO SyGreen Mix displays earlier Ct, cleaner melt peaks and better efficiency compared to each of the competitor mixes.


ROX (6-carboxy-X-rhodamine) is used as a passive reference dye in ROX-dependent real-time PCR instruments to normalize for variations of fluorescence levels that can arise mainly due to optical path variations among wells. Normalisation of the fluorescence intensity (Rn) is done in real-time PCR software by dividing the emission intensity of the specific signal by the emission intensity of ROX.

ROX does not take part in the PCR reaction and its fluorescence levels are not proportional to the quantity of DNA in each well, so the addition of this fluorophore to a mix provides a constant fluorescent signal during amplification.

Different types of real-time PCR instruments requiring a passive reference standard have different optimal concentrations of ROX, mainly due to the different optical configurations of each system (i.e. the different type of excitation source and optics used).

The addition of either too little or too much ROX would result in a very noisy signal impacting on the results of the reaction. Therefore, it is extremely important for the user to:

  • Determine the correct ROX concentration to optimise real-time PCR results, and
  • Check the ROX settings on the software used to set up the reaction

ROX concentrations in qPCRBIO SyGreen Mixes:

The qPCRBIO SyGreen Mixes that contain passive reference dyes come in different formulations, each with a different concentration of the passive reference dye:

  • qPCRBIO SyGreen Mix Lo-ROX (PB20.11) contains 112 nM ROX
  • qPCRBIO SyGreen Mix Hi-ROX (PB20.12) contains 1.12 µM ROX
  • qPCRBIO SyGreen Mix Fluorescein (PB20.13) contains 0.2 mM fluorescein
  • qPCRBIO SyGreen Mix Separate-ROX (PB20.14) 2x mix contains no ROX and include a separate tube of 50 µM ROX additive. This enables you to choose what concentration of ROX you’d like to use.

Available sizes:

PB20.12-01 100 x 20μL Reactions
PB20.12-05 500 x 20μL Reactions
PB20.12-20 2000 x 20μL Reactions
PB20.12-50 5000 x 20μL Reactions (1 x 50mL Bottle)
PB20.12-51 5000 x 20μL Reactions (50 x 1mL in Pouch)

“We’ve found PCR Biosystems’ qPCR master mixes are not only the most competitive on price, but they also give the best results. The introduction of their easy-to-load blue mixes has made an excellent product even better. If your research relies on reliable qPCR analysis then I would look no further than PCR Biosystems. I really can’t recommend their products highly enough.”

Dr Will Cawthorn, University of Edinburgh


PCR Biosystems
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