Reaction Setup
Before starting the reaction setup, thaw LabQ RealTime PCR Kit SYBR Universal and mix thoroughly but gently to ensure even distribution of components.
Dilute your standard DNA and experimental samples with nuclease-free water to the desired concentrations and add them to theier designated wells in the multi-well plate.
For negative control, add nuclease-free water.
Keep the plate on ice until further use.
Component | Volume | Final Concentration |
LabQ RealTime PCR Kit SYBR Universal | 10 µl | 1x |
Template DNA | < 10 ng for low complexity templates | |
X µl | >1 for high complexity templates | |
Max. 2µl of reverse transcription reaction | ||
Forward primer (10µM) | 0.4 µl | 0.05 – 0.09 µM each |
Reverse primer (10µM) | 0.4 µl | 0.05 – 0.9 µM each |
Nuclease-free dH2O | To 20 µl |
Recommended qPCR Protocol
Set | Cycles | °C | Time |
Initial Denaturation | 1 | 95°C | 5 minutes |
Amplification | 40 | 95°C
55-60° 72°C |
10 seconds
15 seconds / 10 seconds 20 seconds |
Add an addtional melting curve step if required. A melting curve is recommended to ensure specific amplification and to detect possible primer oligomers.