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FAQs
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qPCRBIO SyGreen® Chameleon Kit FAQs
Can I handle UNG kits the same way as a standard kit?
No. UNG is more temperature-sensitive than DNA polymerases and should be kept cool (on ice or cooling blocks) during reaction setup. The 20x UNG Additive should also be kept cold at all times with handling limited to the minimum necessary for thawing and reaction setup. An additional UNG activity step is also included in the qPCR thermal cycling program for UNG kits, to allow this enzyme to eliminate any previously amplified uracil-containing DNA.
Does the colour change affect qPCR performance?
Neither the blue dye or SampleTrack Buffer, when used at the recommended concentration, inhibit PCR/qPCR reactions or quench qPCRBIO SyGreen® fluorescence. No change to cycling conditions is required when using SampleTrack Buffer as recommended as part of this kit.
How can I use the SampleTrack buffer provided with this kit?
SampleTrack Buffer allows you to confirm that template has been added to each reaction. The template containing SampleTrack Buffer is yellow; when it is added to the blue qPCR master mix in a tube or well, the reaction turns green.
Add SampleTrack Buffer to each template sample before dispensing the template into the qPCR reactions. Do not add SampleTrack Buffer to the reaction master mix or as a separate component after template addition, as this will not allow you to track whether template has been added.
The required concentration of SampleTrack Buffer in the prepared template depends on the volume of template added to each reaction. Prepare sufficient template containing SampleTrack Buffer for all required reactions, ensuring that SampleTrack Buffer will be at a final concentration of 1x in the completed qPCR reaction.
The examples below provide sufficient prepared template for ten 20 µL reactions. This is sufficient for one target gene and one reference gene, each tested using four technical replicates, with additional volume allowed for pipetting.
| Template added per 20 µL reaction |
SampleTrack concentration required in prepared template |
Preparation for 10 reactions |
|---|---|---|
| 1 µL | 20x | 2 µL 100x SampleTrack Buffer + 8 µL template
10 µL total prepared template |
| 2 µL | 10x | 2 µL 100x SampleTrack Buffer + 18 µL template
20 µL total prepared template |
| 3 µL | 6.7x | 2 µL 100x SampleTrack Buffer + 28 µL template
30 µL total prepared template |
| 5 µL | 4x | 2 µL 100x SampleTrack Buffer + 48 µL template
50 µL total prepared template |
Mix the prepared template thoroughly before dispensing it into the qPCR plate. Dispense the template volume specified in the first column into each reaction. Use the same template volume in all reactions prepared from a given sample. If different template volumes are used, prepare each template with the corresponding concentration of SampleTrack Buffer. Scale all preparation volumes proportionally when more or fewer reactions are required.
How does the colour change work?
The qPCRBIO SyGreen® Blue Mix provided as part of this kit is blue, and SampleTrack Buffer is yellow. When a sample containing SampleTrack Buffer is added to the blue mastermix, the reaction turns green, providing an immediate visual indication that sample addition has occurred.
Is this kit suitable for probe-based qPCR?
No. The qPCRBIO SyGreen® Chameleon Kit is a dye-based qPCR product. For probe-based qPCR, use appropriate probe-based ranges such as Clara® Probe Mixes or Clara® Probe 1-Step Mixes. These can be combined with standalone SampleTrack Buffer for similar colour-change benefits.
What is the main benefit/use of the qPCRBIO SyGreen® Chameleon Kit?
The main benefit is visual setup confirmation. The kit helps users track both mastermix and sample addition during qPCR setup, reducing the risk of failed reactions caused by omitted reagents or missed samples.
What type of qPCR is the qPCRBIO SyGreen® Chameleon Kit used for?
The kit is designed for dye-based qPCR using qPPCRBIO SyGreen® detection. It is suitable for applications such as relative gene expression, absolute quantification and 2-step RT-qPCR workflows.
