remain solid up to 90°C. Alternatively, a 2 × Gelrite concentrate
(12–20 g/L) is made in water, melted by boiling, and
added to an equal volume of 2 × concentrated Sulfolobus
medium (Grogan 1989). Calcium (Ca(NO3)2) and magnesium
(MgCl2) are added to a final concentration of 1.5 and 5 mM,
respectively, to stabilize the gel. Before the gel solidifies, ca.
25 mL is poured into standard (90 mm) Petri plates with cams.
After the Gelrite solidifies, plates can be stored at 4°C indefinitely.
Approximately 0.1 mL, from undiluted to 10–3, of
enrichment cultures are spread on Gelrite plates in the presence
or absence of 0.5 mL 0.2% Gelrite dissolved in Sulfolobus
medium. Plates are incubated inverted in airtight moist containers
at 75–80°C for approximately 1 week before colonies
appear (Fig. 1C). Multiple wet paper towels and a 90-mm Petri
dish filled with water at the bottom of a sealable container
(e.g., Tupperware®) is sufficient.
remain solid up to 90°C. Alternatively, a 2 × Gelrite concentrate
(12–20 g/L) is made in water, melted by boiling, and
added to an equal volume of 2 × concentrated Sulfolobus
medium (Grogan 1989). Calcium (Ca(NO3)2) and magnesium
(MgCl2) are added to a final concentration of 1.5 and 5 mM,
respectively, to stabilize the gel. Before the gel solidifies, ca.
25 mL is poured into standard (90 mm) Petri plates with cams.
After the Gelrite solidifies, plates can be stored at 4°C indefinitely.
Approximately 0.1 mL, from undiluted to 10–3, of
enrichment cultures are spread on Gelrite plates in the presence
or absence of 0.5 mL 0.2% Gelrite dissolved in Sulfolobus
medium. Plates are incubated inverted in airtight moist containers
at 75–80°C for approximately 1 week before colonies
appear (Fig. 1C). Multiple wet paper towels and a 90-mm Petri
dish filled with water at the bottom of a sealable container
(e.g., Tupperware®) is sufficient.
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