Abstract
The protocol describes the procedure of total RNA isolation from cells of the cyanobacterium Synechocystis sp. PCC 6803. This protocol is also applicable to Synechococcus elongatus PCC 7942 and PCC 6301, Thermosynechococcus vulcanus, and other unicellular and filamentous species of cyanobacteria that do not have thick polysaccharide-containing outer layers. For the latter, Trizol-containing protocols should be adapted. The yield of RNA depends on optical density of cyanobacterial culture and may reach up to 10-20 µg of total RNA per 1 ml of cell culture. RNA isolated by this method can be used for Northern blot hybridization, RT-qPCR, microarrays and Next Generation Sequencing.
Keywords: Synechocystis, Cyanobacteria, RNA isolation, RNA quality, Hot phenol
Materials and Reagents
Equipment
Procedure
Notes
Solution of LiCl contains DNA, and it’s important to remove supernatant completely. Also DNA in LiCl solution can be precipitated by 2 volumes of ethanol or 0.8 volumes of 2-isopropyl alcohol (Lab Scan, C19C11X).
Recipes
Acknowledgments
This work was supported by a grant from Russian Science Foundation No. 14-24-00020 to D.A.L. and by a grant from Russian Foundation for Basic Research No. 14-94-01446a to K.S.M.
References
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Hello, pellet may be invisible. Also this protocol is not recommended for DNA isolation
Thanks for prompt reply. And i have quires about growing Synechoscystis in lab. could you assist me on the same. will u calrify it?
Hi, Tarkeshwar, I have never tried to use Trizol for RNA isolation from cyanobacteria