Revision as of 19:50, 26 June 2012

Transformation Synechocystis 6803

Make sure the cells are not in stationary phase. (dark green and bubbles all over)
Take 15 ml of cell culture and spin it down for 15 minutes at RT. (6000xg)
ReSuspend the pellet to 1ml in BG11 broth.
Mix in 500 ng plasmid with 500ul of the 1ml resuspended pellet
Incubate for 5 h at 25 degrees C and in the light.
Plate the cells onto BG11 plates with antibiotic.

Spray your gloves, containers, and working surfaces with ethanol, and let the ethanol dry, before handling cyanobacteria.

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 =Transformation Synechocystis 6803=
-. Make sure the cells are in exponential growth phase. (OD730 of 0.8)
+#Make sure the cells are not in stationary phase. (dark green and bubbles all over)
+#Take 15 ml of cell culture and spin it down for 15 minutes at RT. (6000xg)
+#ReSuspend the pellet to 1ml in BG11 broth.
+#Mix in 500 ng plasmid with 500ul of the 1ml resuspended pellet
+#Incubate for 5 h at 25 degrees C and in the light.
+#Plate the cells onto BG11 plates with antibiotic.
-. Take 15 ml of cell culture and spin it down for 15 minutes at RT.
-. ReSuspend the pellet to 1ml in BG11 broth.
-. Mix in the plasmid to a final concentration of 150 ng/ml
-. Incubate for 5 h at 25 degrees C and in the light.
-. Plate the cells onto BG11 plates with antibiotic.
 Spray your gloves, containers, and working surfaces with ethanol, and let the ethanol dry, before