We have validated three thermocyclers with melt curve capabilities for use with the SALSA® MC002 SMA Newborn Screen. Other thermocyclers may be suitable provided that they meet a number of criteria (see below).
The thermocyclers validated by us are:
- Bio-Rad CFX96 Touch, with Bio-Rad HSP9655 white/white plates and Bio-Rad MSB1001 MicroSeal Plate Sealing film.
- Roche LightCycler 480 I, with BIOplastics B17489 plates and BIOplastics 157300 Opti-Seal adhesive seals.
- Thermo Fisher QuantStudio 5, with Thermo Fisher N8010560 MicroAmp Optical 96 well plates and Thermo Fisher 4360954 MicroAmp Optical Adhesive Film.
Instructions on how to set up these thermocyclers can be found in the instructions for use.
Other thermocyclers should at least meet the following requirements:
- Presence of melt curve capabilities with temperature ranges as specified in the thermocycler program in the instructions for use.
- Ability to detect fluorescence from the Cy5 fluorophore.
- Presence of a heated lid (99–105°C).
Note
Use of a thermocycler not validated at MRC Holland requires in-house validation.