An anaerobic, intrachamber incubator for growth of Methanosarcina spp. on methanol-containing solid media

Abstract

To simplify the incubation of Methanosarcina spp. on solid agar medium, a two-port, manual, rectangular air lock was modified to serve as an anaerobic incubator. In one operation, it is possible to incubate 153 petri plates, the equivalent of 11 standard anaerobic jars, with plating efficiencies identical to those of traditional protocols.

FIG. 1

Line drawing of the air lock body, showing the acrylic panel in place and the three-way valve with gas mixture supply lines.

FIG. 2

(A) Modified air lock-incubator with the door partially open before installation inside an anaerobic chamber. The metal housing of the door frame (a), slotted bracket (b), and U-shaped bracket and pin holding a swivel threaded-bolt with a threaded knob on the free end (c) are indicated. (B) Incubator with door locked and tightened against black rubber seal by knobs on threaded swivel bolts. The rubber vacuum hose for attachment to anaerobic jar (d) is indicated. (C) Incubator installed inside an anaerobic chamber. (D) Space fillers for use inside incubator.