Boost your qPCR results without inhibition with GreenDye!
GreenDye is a highly sensitive DNA binding dye suitable for qPCR applications and melting curve analysis.
GreenDye shows very low DNA polymerase inhibition when compared with other dyes in PCR. It features a very high fluorescence when bound to double-stranded DNA and exhibits no degradation during PCR cycling. It is a perfect dye for qPCR and high resolution melting applications.
For research use and further manufacturing.
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- real-time PCR (FAM channel)
- melt curve analysis
- High Resolution Melting (HRM)
- real-time primer extension
It is recommended to use GreenDye at 1x concentration.
For example, add 1.25 μl GreenDye to a 25 µl PCR reaction. The fluorescence is measured at the usual SYBR/FAM channels.
GreenDye 20x is tested together with HiDi DNA polymerase (#9001) in a real-time PCR. A sequence in genomic HeLa DNA is used as a target to validate ct- and fluorescence values of GreenDye at 1x concentration.
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No contamination has been detected in standard test reactions.
The fluorescence of GreenDye is benchmarked using a qPCR assay for the detection of a sequence in genomic HeLa DNA. PCRs were performed with 1 ng/µl of HeLa gDNA in the presence of either GreenDye 1x, EvaGreen® 1x or SYBR® Green I 1x in combination with HiDi DNA polymerase.
The real-time PCR curve with 1x GreenDye (green) in comparison with 1x EvaGreen (grey) and 1x SYBR® Green I (blue) shows the lowest ct-value.
The melting curves are acquired from the PCR experiment described above. All three dyes show a single peak indicating a single amplification product. GreenDye however, shows the sharpest melting peak.