qPCRBIO SyGreen Mixes – Witec AG

The 2x mix uses a proprietary DNA 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.

qPCRBIO SyGreen® Blue Mix contains a non-reactive dye to improve reaction mix visibility and enable sample visualisation in reaction plate wells, allowing greater pipetting precision and reduced errors without affecting your real-time PCR performance.

Applications

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

SYBR Green vs SyGreen, do they perform the same way?

Yes, SyGreen and SYBR Green (registered trademark of Molecular Probes Inc. and is owned by Thermo Fischer Scientific) use the same molecular mechanism to generate signal in qPCR and SyGreen reagents are an alternative to SYBR Green-containing master mixes.

How does qPCRBIO SyGreen® work?

These intercalating dyes possess aromatic and planar groups that allow the intercalation of the molecule in the minor groove of double-stranded DNA spanning approximately 3.5-4 nucleotides. The positive groups present stabilise the dyes allowing for the coordination with the negatively charged DNA backbone.

Due to this mechanism of binding to DNA, they are not able to bind to single-strand nucleic acids, unless secondary structures are formed. On the other hand, fluorescence increases up to 1,000-fold upon intercalation with dsDNA.

When a dye intercalates between DNA base pairs, it emits fluorescence proportional to the amount of bound dye (and hence to the amount of DNA in a sample). This allows visualisation of nucleic acids on gels or fluorimeters in real time qPCR instruments. In the latter case, the number of intercalated dye molecules in a sample increases with the progress of a qPCR, leading to a corresponding increase in fluorescence with each cycle. At the onset of the reaction, the free dye emits a basal fluorescence level. As the reaction proceeds the fluorescence intensity increases exponentially, before it plateaus in later cycles, leading to the classic sigmoidal amplification plot.

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.