Bandit Biosystems panels are built for open qPCR instruments that labs already own. Most of our multiplex panels use FAM, HEX, Cy5 and ROX. These tips cover the setup questions we hear most often.
Before the first run
- Check your channels. Confirm that your instrument reads all four dyes the panel uses. HEX can be read as VIC or JOE where there is no HEX channel.
- Check dye calibration. Use calibrations for the dyes you will run, as your instrument's manufacturer recommends.
- Use the product page. Each product page lists the tube and dye layout, reaction setup and recommended cycling conditions.
Instrument notes
- Applied Biosystems QuantStudio and 7500 series. These instruments often use ROX as a passive reference dye. When ROX is a reporter dye in the panel, set the passive reference to None so the ROX target signal isn't used for normalization.
- Roche LightCycler 480. Multiplex runs typically need a color compensation file to correct for overlap between channels. Make one for the dye set before reporting multiplex results.
- Bio-Rad CFX96, CFX384 and CFX Opus. Assign the panel's fluorophores to each well in the plate setup so every channel is collected and analyzed.
During analysis
- Set thresholds per channel rather than one threshold for all dyes.
- Review the positive and no-template controls before reading sample wells.
- Check the internal control in every sample well.
If you'd like a plate template or help with a new instrument, contact our applications team.