Materials and Reagents

1. Anabaena sp. PCC 7120
   
2. BG11₀ growth medium-BG11 medium without addition of combined nitrogen (Rippka et al., 1979)
   
3. 1 M 2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]ethanesulfonic acid-KOH (TES) (pH 8.2)
   
4. 1 M 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid-NaOH (Hepes) (pH 7.2)
   
5. 1 M NaCl
   
6. 1 M Na-EDTA
   
7. Sucrose (Merck KGaA, catalog number: K45136551417 )
   
8. Lysozyme (N-acetylmuramide glycanhydrolase) (Roche Diagnostics, catalog number: 10837059001 )
   
9. Extraction buffer (see Recipes)
   

Equipment

1. 50 ml Falcon tubes
   
2. 500 ml centrifuge vials (optional)
   
3. Spectrophotometer (e.g. PerkinElmer, model: Lambda 25 )
   
4. Sorvall GS-3 rotor centrifuge for 500 ml vials (optional)
   
5. SL 16R centrifuge (6 x 50 ml) (e.g. Thermo Fisher Scientific)
   
6. Vortex
   
7. Sonicator (e.g. Labsonic U, B. Braun Melsungen AG)
   
8. Orbital shaker (e.g. Infors HT, Labotron)
   
9. Incubator (e.g. Certomat, B. Braun Melsungen AG)
   
10. Bright-field microscope (40x magnification)
    
11. Water bath or thermostat at 37 °C for Chlorophyll measurements
    

Procedure