Kit Components of RealStar® PCR Kits (RUO/CE)
All RealStar® PCR Kits are ready to use real-time PCR systems which consist of master reagents containing enzyme, buffer, primer and probes as well as an internal control, a positive control or quantification standards and PCR grade water.
- Complete, ready to use real-time PCR reagents systems and kits
- Available for most common real-time PCR instruments
- Validated with a number of extraction systems
Master A and Master B contain all components (PCR buffer, DNA polymerase, magnesium salt, primers and probes) to allow PCR mediated amplification and detection of pathogen specific DNA/RNA and Internal Control in one reaction setup.
- Single pipetting step
- 12 rxn per tube
All RealStar® PCR Kits (CE) and all RealStar® reagent systems (RUO) contain a heterologous Internal Control (IC), which can either be used as a PCR inhibition control or as a control of the sample preparation procedure (nucleic acid extraction) and as a PCR inhibition control.
- Non-related (randomized) sequence
- Amplification with specific primer
- Detection with specific probes
- Non-competitive amplification – analytical target with highest sensitivity
- High positive samples may lead to a reduction (low fluorescence) of Internal Control signal
If the IC is used as a PCR inhibition control, but not as a control for the sample preparation procedure, set up the Master Mix according to the following pipetting scheme:
If the IC is used as a control for the sample preparation procedure and as a PCR inhibition control, add the IC during the nucleic acid extraction procedure.
The IC should always be added to the specimen/lysis buffer mixture. The volume of the IC which has to be added, always and only depends on the elution volume. It represents 10% of the elution volume.
If the IC was added during the sample preparation procedure, set up the
Master Mix according to the following pipetting scheme:
The Quantification Standards contain standardized concentrations of specific DNA/RNA and can be used individually as positive controls, or together to generate a Standard curve, which can be used to determine the concentration of specific DNA/RNA in a sample.
- External standards
- Generation of standard curve
- Copies/ml or IU/ml
- Purified RNA/DNA
- Calculation of original viral load in sample (ml)