Question: Which thermal cyclers are compatible with the Fixed RNA Profiling assay?
Answer: Listed below are the minimum thermal cycler specifications for compatibility with the Fixed RNA Profiling Assay:
- Uniform heating of 100 μl emulsion volumes.
- The thermal cycler supports uniform temperature cycling.
- The thermal cycler has a temperature-controlled lid.
Please check with the thermal cycler manufacturer to ensure the above specifications are met.
The following thermal cyclers have been validated to be compatible with the Fixed RNA Profiling Assay:
- Bio-Rad C1000 Touch™ Thermal Cycler with 96-Deep Well Reaction Module* (#1851197)
- Eppendorf MasterCycler® X50S (Cat No. North America 6311000010)
- Biometra TAdvanced 96SG Endpoint Thermal Cycler (#846-4-070-241)
If using thermal cyclers other than BioRad C1000, the ramp rates should be adjusted for all the steps as described below l Eppendorf MasterCycler X50s: 3°C/sec heating and 2°C/sec cooling l Analytik Jena Biometra TAdvanced: 2°C/sec heating and cooling.
Limited testing in-house has suggest that assay performance (i.e. decrease in library complexity) may be affected if an alternative, not validated thermal cycler is used. This may be exacerbated by the high cell loads in this assay. The Veriti 96-Well Thermal Cycler (4375786) from Thermal Fisher Scientific did not make the final list of validated thermal cyclers for Fixed RNA Profiling assay.
Products: Fixed RNA Profiling