FAQs
Get the most out of your enzyme with our documents, guides, brochures and tools
FAQs
Browse our frequently asked technical questions by selecting a product below.
How can I use SampleTrack Buffer with PCRBIO reagents?
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, when added to purple mixes reactions turn orange, when added to red mixes (at 5x final concentration) reactions turn bright red – orange, and when used with clear mixes these become yellow.
Add SampleTrack Buffer to each template sample before dispensing the template into the PCR or 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 or PCR reaction for clear, blue and purple mixes and at 5x concentration in red endpoint mixes.
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. Adjust volumes accordingly for 25, and 50 μL reactions, depending on your application and which PCR Biosystems reagent you are using.
| Template added per 20 µL reaction |
Concentration required in template for 1x final concentration |
Preparation for 10 reactions at 1x final concentration |
Concentration required in template for 5x final concentration |
Preparation for 10 PCRBIO Red Mix reactions at 5x final concentration |
|---|---|---|---|---|
| 1 µL | 20x | 2 µL 100x SampleTrack Buffer + 8 µL template 10 µL total prepared template |
100x | Not applicable Each reaction requires 1 µL of 100x SampleTrack Buffer, leaving no volume for template + sample in 1 µL. |
| 2 µL | 10x | 2 µL 100x SampleTrack Buffer + 18 µL template 20 µL total prepared template |
50x | 10 µL 100x SampleTrack Buffer + 10 µL template 20 µL total prepared template |
| 3 µL | 6.7x | 2 µL 100x SampleTrack Buffer + 28 µL template 30 µL total prepared template |
33.3x | 10 µL 100x SampleTrack Buffer + 20 µL template 30 µL total prepared template |
| 5 µL | 4x | 2 µL 100x SampleTrack Buffer + 48 µL template 50 µL total prepared template |
20x | 10 µL 100x SampleTrack Buffer + 40 µL template 50 µL total prepared template |
Mix the prepared template thoroughly before dispensing it into the PCR plate or wells. 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.
