1X Tris-EDTA buffer (TE), autoclaved and stored at room temperature. We use a 100X stock solution (VWR International, USA), diluted to 1X in ultrapure water (Milli-Q, Merck, DE).
SYBR Green I nucleic acid gel stain (ref. S9430, Merck). We use aliquots diluted to 100X in autoclaved TE and stored at -20°C.
Dry bath with heat block for 96 wells and heated lid, or thermal cycler with heated lid. The heated lid reduces condensation, and this helps prevent contamination and sample loss. We use a ThermoFisher Compact Digital Dry Bath S (ref. 88871003, ThermoFisher, USA) with 96-well block (ref. 88871107). Instead of a heated lid, we use four ‘6x1.5ml’ blocks (ref. 88871103) which we heat and place on top of the plate (see below).
An appropriate 96-well PCR plate and sealing film for use in the dry bath or thermal cycler. The plate should fit the 96 well block perfectly: there should be no air between the wells and the heating element, and the wells should not extend above the heating element. We have also found that the brand of the plate affects the precision of the results. We use low profile PCR plates from Starlab (ref. E1403-0200, Starlab, DE) and adhesive PCR film (ref. 732-3212, VWR).
An appropriate 96-well plate for use in the flow cytometer, and a seal for this plate. We use Rotilabo microtitration plates (ref. 9291.1, Carl Roth, DE) and aluminum foil seals (ref. 391-1282, VWR).
A flow cytometer that can record the analyzed volume, and ideally with an automated loader. We use a BD FACSVerse (BD Biosciences, USA).
Vortexer and plate centrifuge.
Multichannel pipettes are highly recommended, particularly when measuring samples stored in a 96-well plate.