qPCRBIO SyGreen Blue Mix Hi-ROX – Witec AG

Art No | PB20.16
from 53.00 CHF

qPCRBIO SyGreen Blue Mix is particularly useful in reducing errors when loading small volume reactions or using white plates.


This easy-to-see mix uses a non-reactive blue dye for easy sample visualisation during reaction setup, allowing fast, highly sensitive and reproducible real-time PCR with more precision and accuracy than ever.

  • Easy-to-see blue mix for greater pipetting precision and accuracy
  • 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
  • Compatible with all standard and fast cycling real-time instruments

The blue mixes contain a non-reactive dye to improve reaction mix visibility, allowing greater pipetting precision and reduced errors without affecting your real-time PCR performance. qPCRBIO SyGreen Blue Mix can be used to quantify any DNA template including genomic, cDNA and viral sequences, and is able to reliably detect extremely low copy number targets. The antibody-mediated hot start system prevents the formation of primer dimers and non-specific products leading to greatly improved reaction sensitivity and specificity.

This 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 Blue 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 Blue Mixes:

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

  • qPCRBIO SyGreen Blue Mix Lo-ROX (PB20.15)  contains 112 nM ROX
  • qPCRBIO SyGreen Blue Mix Hi-ROX (PB20.16) contains 1.12 µM ROX
  • PCRBIO SyGreen Blue Mix Separate-ROX (PB20.17) 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:

PB20.16-01 100 x 20μL Reactions
PB20.16-05 500 x 20μL Reactions
PB20.16-20 2000 x 20μL Reactions
PB20.16-50 5000 x 20μL Reactions (1 x 50mL Botle)
PB20.16-51 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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