Culturing Chlamydomonas and CC4533 Strains

Chlamydomonas wild type strain 4A+ (CC- 4051 4A+ mt+) was maintained in the lab on TAP agar medium in dim light intensities (15-20 µmol m
^-2s
^-1) at 22°C (room temperature). Standard TAP medium recipe can be found at the website of
Chlamydomonas Resource Center. Hutner’s trace element solution is an ingredient in the TAP medium. Hutner’s trace element recipe can also be found on the
Chlamydomonas Resource Center website. Our lab’s TAP medium recipe is slightly different from the standard one and can be found at
https://doi.org/10.17504/protocols.io.bgzujx6w^{41 (link)}. Liquid 4A+ cultures were grown in liquid TAP medium under low light (50-60 µmol m
^-2s
^-1) for 3 days on a New Brunswick Scientific Excella E5 platform shaker (Enfield, CT) at 150 rpm for aeration. CC4533 bacterial strain stock was maintained in the lab under dim light (15-20 µmol m
^-2s
^-1) at 22°C on either TAP or LB agar medium. Liquid cultures of CC4533 were grown in culture tubes in 3 mL of TAP medium at 22°C on a MaxQ420HP incubator shaker (Thermo Fisher Scientific, Waltham, MA) at 200 rpm for aeration. Light intensities were measured using a LI-250A light meter (LI-COR, Inc., Lincoln, NE).