A multiplex qPCR result is only as useful as the controls around it. Our panels use three kinds, and each answers a different question.
Internal controls: did this sample work?
An internal control is amplified in the same well as the targets. If it comes up, the extraction recovered nucleic acid and the reaction was not inhibited.
- Human RNase P (hRPP) is used in our human-specimen panels. Because it is a human gene, it also indicates that the sample contained human material.
- SPRY1 is the multi-species internal control in our veterinary panels, chosen to work across specimens from different animal sources.
A sample whose internal control fails to amplify, with no targets detected, cannot be called negative. It points to a problem with extraction, inhibition or sample quality.
Positive controls: did the run work?
Every Bandit Biosystems panel includes panel-matched positive controls. They contain template for the panel's targets and should give a signal in every expected channel. Running them on each plate confirms that the primer and probe mixes, master mix, cycling program and instrument channels performed as expected.
Some kits offer a choice. Women's Health Panel C Ready-2-Test, for example, comes either with four individual positive controls or with a single Super Control covering all targets in one tube.
No-template controls: is anything contaminated?
A no-template control replaces sample with nuclease-free water. It should show no amplification. A signal in a no-template well points to contamination of reagents, pipettes or the work area.
Setting acceptance criteria
Each laboratory sets its own acceptance criteria, such as Cq cut-offs for controls and how to handle a failed internal control, during its own verification. Our Field Applications Specialists can help you plan that work.