Table 1.

Protocols tested for assessing inactivation using lysis buffers.

Manufacturer, RNA extraction kit, platform	Reagents (volume/sample)	Active virucidal components*	Reagent:sample ratio
Qiagen,	ACL buffer (190 µl)	GITC 30% to <50%	1.6:1
QIAamp 96 Virus QIAcube HT Kit
(Cat #: 57731),	ATL buffer (100 µl)	1% to <3% SDS
Qiagen QIAcube HT.
(Referred to here as Qiagen protocol)	Proteinase K (20 µl)
	Carrier RNA (5 µl)
	MS2 (10 µl)
Thermo Fisher,	Lysis binding buffer (350 µl)	GITC 55%–80% <0.001% acrylamide	3.8:1
MagMAX Pathogen RNA/DNA kit		Zwittergent
(Cat #: 4462359),
KingFisher Flex.
(Referred to here as MagMAX Protocol 1)	Isopropanol (300 µl)	100% 2-propanol
	Carrier RNA (2 µl)
	Water (100 µl)
	MS2 (10 µl)
Thermo Fisher,	Lysis binding buffer (265 µl)	GITC 55%–80% <0.001% acrylamide	1.4:1
MagMAX viral/pathogen nucleic acid isolation kit		Zwittergent
(Cat #: A48310),
KingFisher Flex.
(Referred to here as MagMAX Protocol 2)	Proteinase K (5 µl)
	†Water (magnetic beads) (10 µl)
	MS2 (10 µl)

^*As identified directly from components or manufacturer information or inferred from the associated MSDS.

^†Water was used to replace the magnetic beads, as the washing steps described below would not remove the beads, and the beads interfered with the read-out of the TCID₅₀ assay.

GITC, Guanidinium thiocyanate; SDS, Sodium dodecyl sulphate.