Table 4.

GT RT-qPCR test kit formulation

Component	Stock	Volume (μl)	Final concentration
Template	Quantitative Synthetic SARS-CoV-2 RNA: ORF, E, N (ATCC® VR-3276SD™)	0.5–5	5–50,000 copies
	Full-length viral RNA + HEK293T total RNA	5	5–50,000 copies viral RNA + 0.02-200 ng of HEK293T total RNA
Primer/probe:	GT singleplex or multiplex primer/probe mix	1.5	500 nm primers, 125 nm probes
GT-Master Mix1	5× buffer:     250 mm Tris, pH 8.3,     375 mm KCl,     15 mm MgCl2,     47.5% trehalose	4.0	1× buffer:     50 mm Tris, pH 8.3,     75 mm KCl,     3 mm MgCl2,     9.5% trehalose
	10 mm dNTP	0.8	400 µm
	100 mm DTT	1.0	5 mm
	GT-rRI2	1.0	50 µg/ml
	GT-His-Taq2	1.0	7.5 µg/ml
	GT-MMLV2	0.5	3.3 µg/ml
	20 mg/ml BSA	1.0	1 mg/ml
	25 µM ROX	0.4	500 nm
Water	Molecular biology grade water
	Total volume	20

¹This can be prepared as a 2× master mix and stored at −20 °C.

²See “Experimental procedures.” Concentration or volume should be adjusted for activity.