IGEM:Harvard/2006/Cyanobacteria/Protocols: Difference between revisions
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Revision as of 13:35, 22 June 2006
Media
1L agar for freshwater cyanobacteria (no glucose)
This protocol produces 1L agar suitable for freshwater cyanobacteria (PCC7942 and PCC6803). [1]
- Mix 10 g of agar and 1 mL thiolsulfate, top off with H20 to 500 mL total volume.
- Autoclave the product of (1)
- Mix 20mL of 50x BG-11 solution and 480 mL of H20
- Autoclave the product of (3)
- Mix (2) and (4), pour plates and let cool
1L agar for freshwater cyanobacteria (with glucose)
This protocol produces 1L agar with 5 mM glucose suitable for PCC6803 cyanobacteria.
- Mix 10 g of agar and 1mL thiolsulfate, top off with H20 to 500 mL total volume
- Autoclave the product of (1)
- Mix 20 mL of 50x Sigma BG-11 solution and 230 mL of H20
- Autoclave the product of (3)
- Mix 5 mL glucose and 245 mL of H20 [2]
- Autoclave the product of (5)
- Mix (2) and (4) and (6), pour plates and let cool
References
[1]: The use of thiosulfate is discussed in the following papers; it is isn't required, but it supposedly improves growth rate.
- Thiel T, Bramble J, and Rogers S. Optimum conditions for growth of cyanobacteria on solid media. FEMS Microbiol Lett. 1989 Oct 1;52(1-2):27-31. DOI:10.1016/0378-1097(89)90164-x |
[2]: Usage of glucose is to take advantage of heterotrophic growth of PCC6803 with glucose; this supposedly cuts down the growth cycle to 4 days, according to the 2006 MIT iGEM team.
- Spence E, Bailey S, Nenninger A, Møller SG, and Robinson C. A homolog of Albino3/OxaI is essential for thylakoid biogenesis in the cyanobacterium Synechocystis sp. PCC6803. J Biol Chem. 2004 Dec 31;279(53):55792-800. DOI:10.1074/jbc.M411041200 |