Evaluated addition of fluorescein to Synechocystis cells for standardizing 77K fluorescence spectra (excited at 435 nm). Measured absorption at peak 490nm: 0.53 stock solution was diluted 1:3 -> corrected absorption for stock solution at peak 2.12 from: https://en.wikipedia.org/wiki/Fluorescein Extinction coefficient at 490 nm = 76900 Mol-1 cm-1 Stock solution concentration: A=e*L*C C=A/(e*L) C=2.12 / (76900 cm-1 M * 1 cm…
Christopher Williams joined the lab
March 10, 2012 Chris Williams has started working in the lab. Chris will investigate hydrogen metabolism in the cyanobacteria Synechocystis sp. PCC 6803 during his Master’s thesis.
Absorption and Fluorescence
Jablonski diagram Jablonski diagram representing energy levels and spectra. Solid arrows indicate radiative transitions as occurring by absorption (violet, blue) or emission (green for fluorescence; red for phosphorescence) of a photon. Dashed arrows represent non-radiative transitions (violet, blue, green, red). Internal conversion is a non-radiative transition, which occurs when a vibrational state of a higher electronic state is coupled to…
Absorption & Fluorescence spectra Synecho
Whole Cell Absorption 77K [wpfilebase tag=file id=330 tpl=simple /] Thylakoid and purified PSII 77K fluorescence [wpfilebase tag=file id=329 tpl=simple /] A: Excitation at 600 nm excites the phycobilisome and preferentially PSII B: Excitation at 420 nm excites the chlorophylls contained in PSI and PSII PSI , PSI-IsiA and IsiA fluorescence at 77K [wpfilebase tag=file id=328 tpl=simple /]
Sum-up of pre-experiments – Anaerobic incubation of Nannochloropsis
The experiment on anaerobic incubation of Nannochloropsis will determine what anaerobic incubation does to photosynthesis, and to the production of photosynthetic lipids, storage lipids, and ω-3 fatty acids. The pre-experiment was performed in order to find out what would be the ideal time point for making the cells anaerobic. The cells were grown in the same conditions as will be used…
Chloroplast transformation
Potential genes for deletion These genes are not essential components of PSII in Synechocystis psbX psbY psbZ These genes are involved in the light-independent chlorophyll synthesis chlL chlN [wpfilebase tag=file id=333 tpl=simple /] information concerning light-dependent and light independent chlorophyll synthesis Potential neutral region in chloroplast genome The region is downstream of atpB in the Chlamydomonas chloroplast genome (see Purton(2007)_ToolsTechChloroTrans).…
Cryopreservation of marine algae, Nannochloropsis
1 cryopreservation Cultivate N. oceanica or N. gaditana on a shaker (150 rpm, 23°C, 100 yE) to a cell concentration of 1*107 cells/ml. Place culture for 20 hours in the dark before cryopreservation. Equilibrate the CoolCell freezing container (check if black core is inside) at 4°C. Prepare a sterile filtered 20% solution of DMSO in the culture medium used to grow the algae. harvest the…
Expression of genes in the chloroplast
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Nannochloropsis Antibiotics
These antibiotics have been evaluated as selection markers. The used concentration is shown in brackets. Evaluated by: [wpfilebase tag=file id=130 tpl=simple /] [wpfilebase tag=file id=134 tpl=simple /] zeocin (2 μg / mL) blasticidin S (50 μg / mL) hygromycin B (300 μg / mL) Evaluated by: [wpfilebase tag=file id=131 tpl=simple /] zeocin (3 μg / mL) Evaluated by: [wpfilebase…