Chlamydomonas reinhardtii
Chlamydomonas reinhardtii, Mix-lowZnFeCuO2-NO3-MM
strain
CC124(137c) and CC1021(2137)
in vivo conditions
Cells were incubated in the light in Zn-free or Fe-limited TAP, or TAP with NO3 instead of NH4, or in Cu-free minimal medium; for anaerobiosis, cells were incubated in sealed flasks in the dark in TAP
library_type
RAPID
library_description
"Minus Co/Zn: C. reinhardtii strain CC-1021 cells were grown in modified TAP medium with no Co and no Zn at 24 Celsius, 180 RPM shaking, and under continuous light at 120 µmol/m^2*s. RNA was isolated from cells that were collected at a cell density of 2.9x10^6 cells/ml. Fe 0.25ÂµM: C. reinhardtii strain CC-1021 cells were grown in modified TAP medium with 0.25 ÂµM Fe-EDTA at 24 Celsius, 180 RPM shaking, and under continuous light at 90 Âµmol/m^2*s. Cells were transferred to 1ÂµM Fe medium twice prior to experimentation and inoculated in 0.25 ÂµM Fe medium at 1x10^4 cells/ml. RNA was isolated from cells collected at a cell density of 3.0x10^6 cells/ml. Fe 1ÂµM: C. reinhardtii strain CC-1021 cells were grown in modified TAP medium with 1 ÂµM Fe-EDTA at 24 Celsius, 180 RPM shaking, and under continuous light at 90 umol/m^2*s. Cells were transferred to 1ÂµM Fe medium twice prior to experimentation and inoculated in 1 ÂµM Fe medium at 1x10^4 cells/ml. RNA was isolated from cells collected at a cell density of 3.0x10^6 cells/ml. Dark 30 min: C. reinhardtii strain CC-124 cells were grown in TAP medium at 24 Celsius, 140 RPM shaking, and under continuous light at 80 Âµmol/m^2*s. Cells at 4x10^6 cells/ml were transferred to sealed bottles and grown in the dark. RNA was isolated from cells collected after 30 minutes. Dark 2 hrs: C. reinhardtii strain CC-124 cells were grown in TAP medium at 24 Celsius, 140 RPM shaking, and under continuous light at 80 Âµmol/m^2*s. Cells at 4x10^6 cells/ml were transferred to sealed bottles and grown in the dark. RNA was isolated from cells collected after 2 hours. Nitrogen: C. reinhardtii strain CC-1690 cells were grown in TAP medium or TAP -N +KNO3 at 24 Celsius, 140 RPM shaking, and under continuous light at 95 Âµmol/m^2*s. RNA was isolated from cells collected at a cell density of 3.0x10^6 cells/ml. RNA samples from the two nitrogen source samples were pooled to generate one sample. Minus Cu: C. reinhardtii strain CC-1021 cells were grown in modified TAP medium with no Cu at 24 Celsius, 200 RPM shaking, bubbled air, and under continuous light at 90 Âµmol/m^2*s. Cells were transferred to -Cu medium twice prior to experimentation and inoculated at 1x10^4 cells/ml. RNA was isolated from cells collected at a cell density of 3x10^6 cells/ml. The seven RNA samples were pooled in equal proportion for sequencing. Sample provided by Sabeeha Merchant, Anja Hemschemeier, Eugen Urzica, Janette Kropat, Aimee Terauchi and Madeli Castruita."