Isolation of total DNA from Synechocystis sp. PCC 6803 v1

This protocol can be used for extraction of total genomic DNA from Synechocystis sp. PCC 6803. </p

Overlap extension PCR v1

Linear assembly of PCR fragments. Can be used to quickly and efficiently fuse promoters, terminators, fusion proteins etc. without time-consuming sub-cloning steps. </p

Recipe for standard BG-11 media v1

Stanier RY, Deruelles J, Rippka R, Herdman M, Waterbury JB: Generic Assignments, Strain Histories and Properties of Pure Cultures of Cyanobacteria. Microbiology 1979, 111:1–61. Recipes for standard and alternative BG11 for culturing freshwater cyanobacteria, such asSynechocystis sp. PCC 6803, as described. Media is usually not suitable for marine cyanobacteria. </p

Homemade Gibson Mastermix v1

Recipe for homemade 1.33 x Gibson Mastermix. </p

Isolation of plasmid DNA from E. coli (Alkaline lysis method) v1

Isolation of plasmid DNA from E. coli using the alkaline lysis method modified from Birnboim et al., 1979. This protocol is suitable for fast, cheap recovery of large amounts of plasmid, e.g. for cloning purposes or restriction analysis. However, the purity of plasmid is insufficient for sequencing. </p

Recipe for standard BG-11 media v1

Stanier RY, Deruelles J, Rippka R, Herdman M, Waterbury JB: Generic Assignments, Strain Histories and Properties of Pure Cultures of Cyanobacteria. Microbiology 1979, 111:1–61. Recipes for standard and alternative BG11 for culturing freshwater cyanobacteria, such asSynechocystis sp. PCC 6803, as described. Media is usually not suitable for marine cyanobacteria. Final Concentration of Medium. CaCl2*2 H2O 0.036 g/L Citric acid 0.006 g/L NaNO3 1.4958 g/L MgSO4* 7 H2O 0.0749 g/L 0.25M Na2EDTA (pH 8) 0.0056 mL/L Na2CO3 20 µg/L Fe(III) Ammonium citrate 6 µg/L K2HPO4 * 3H2O 30 µg/L TES Buffer (pH 8) 10 mM H3BO3 2.86 mg/L MnCl2 * 4 H2O 1.81 mg/L ZnSO4 * 7 H2O 0.222 mg/L Na2MoO4 * 2 H2O 0.390 mg/L Co(NO3)2 *6 H2O 0.049 mg/L (CuSO4 * 5 H2O 0.079 mg/L if required) </p

Cell lysis and extraction of total protein from Synechocystis sp. PCC 6803 v1

Cell lysis and extraction of total protein. Soluble and insoluble fractions can be further separated. Total protein can be then used for quantification and Western Blot analysis. </p

Isolation of total RNA from Synechocystis (PGTX method) v1

Isolation of 40-200 µg of total RNA from Synechocystis sp. PCC 6803. Can be used for standard applications such as Northern Blot and qPCR. </p

Preparation of chemically competent cells v1

Standard protocol to prepare cells for highly efficient DNA uptake using CaCl2. </p