| Reference |
Zavřel, T., Očenášová, P., Červený, J (2017): Phenotypic characterization of Synechocystis sp. PCC 6803 substrains reveals differences in sensitivity to abiotic stress. PLoS ONE 12(12): 1-21, DOI: 10.1371/journal.pone.0189130PubMed ID29216280
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| Method |
P.7 3rd paragraph: "After harvesting from the photobioreactor, 10 ml of culture suspension was centrifuged (4 000 g, 15 min), supernatant was discarded and pellet was resuspended in 5 ml of distilled water. Aliquots of 480 μl were transferred to eight Eppendorf tubes. The samples were centrifuged (20 000 g, 10 min, 4˚C), supernatant was discarded and 480 μl of cold methanol was added to each tube. The samples were homogenized by gentle pipetting, placed to +4˚C for 20 min and centrifuged again at 4˚C. The
methanol supernatants were used for analysis of chlorophyll a and carotenoids as described above and the pellets were stored in -80˚C until further processing for up to 3 months. After thawing on ice, the pellets were resuspended in 200 μl of 30% KOH and they were incubated at 95˚C for 90 min. After cooling down at laboratory temperature, the samples were mixed
with 1.2 ml of cold ethanol (+4˚C) and stored at -20˚C overnight. The precipitated carbohydrates were then separated by intensive centrifugation (20 000 g, 70 min, 4˚C). The supernatant was discarded and the pellet was dried at 60˚C for 30 min in Vacuum Concentrator 5305
(Eppendorf, Hamburg, DE). Dry pellets were diluted in 480 μl of deionized water and transferred to 96-well plate. Content of carbohydrates in the samples was measured spectrophotometrically by Multiskan™ GO Microplate Spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) at 490 nm, using D-glucose as a standard." |