DNA / PCR

GreenDye 20x

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.

 

Article Product Size Price
 #2000 S GreenDye 20x 200 reactions à 25 µl, 1 x 250 µl  € 50,-
 #2000 M GreenDye 20x 800 reactions à 25 µl, 1 x 1000 µl  € 150,-

Bulk offer: Should you require larger amounts (> 2000 µl), significant discounts are available. Please contact us to receive an individual quotation.

 

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Features

Features

  • Thermally stable for 50 cycles and more
  • Suited for qPCR, melt curve analysis and High Resolution Melting (HRM)
  • No decrease in DNA polymerase performance at 1x concentration
  • Optimized for PCR experiments with HiDi DNA polymerase and HiDi Taq DNA polymerase

Applications

Applications

  • real-time PCR (FAM channel)
  • melt curve analysis
  • High Resolution Melting (HRM)
  • qPCR
  • real-time primer extension
  • screening

Reaction Setup

Reaction Setup

It is recommended to use GreenDye at 1x concentration.
For example, in a 25 µl PCR reaction 1.25 μl GreenDye is used for each reaction. The fluorescence is measured at the usual SYBR/FAM channels.



Example Results

Example Results

 

Benchmarking GreenDye against competitors

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.

 

 

 


GreenDye Melt Curve Analysis

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.
 
 
 
 

 

 

 

Recommendations

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.

 

Content of the Product

20x GreenDye in a aqueous solution

 

Storage

This product is shipped in cool packs. Please store the product upon arrival at - 20 °C. Minimize the number of freeze-thaw cycles by storing in aliquots. For a day-to-day use, we recommend keeping an aliquot at 4 °C.

 

Quality Control

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.
Please inquire more information at info@mypols.de for lot-specific information.
No contamination has been detected in standard test reactions.

 

 

 

img-faqsFAQ`s

Is it possible to use GreenDye in concentrations higher than 1x?

Yes, GreenDye can be used at higher concentrations than 1x. This is accompanied by an increase in fluorescence but may also lead to DNA polymerase inhibition.

Can GreenDye be used in combination with other DNA polymerases than the HiDi DNA polymerase?

Yes, GreenDye is compatible with the majority of DNA polymerases and PCR protocols.


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