I am Trying to grow Synechocystis ATCC 27184 using BG-11 broth. Also providing ligth source 2000-3000 lux using 40Watt LED bulb as light souce. Incubated at 26°C for 1 to 2 weeks and zero hr(initial OD was 0.005). After incubation , Microscopy observation revealed growth of cells with Optical density of 0.68.
But Green color pigmentation was not seen also turbidity also not much. Can anyone tell where i am going wrong????
Initial OD 0.005 is very low number of cells, it would take a lot of time for cells to grow to a visibly turbid amount, this is normal. I would advise you to grow Synechocystis in a smaller volume flasks (50-100 ml) and use that as an inoculum for larger cultures, that is, inoculate to a higher zero hour OD and you will observe reported growth patterns.
The light intensity you use is about right (50 micromole photons per m2 per s or so is good). As Vamsi pointed out, a starting OD730 of 0.005 is very low; I recommend more like 0.1 to be safe. And best to bubble with air; otherwise (on shakers or so) they quickly get carbon-limited. I think that the growth leading to OD 0.68 without green color is due to contamination or so, so good to make sure the strain is axenic.
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