Template:SJTU-BioX-Shanghai/Notebook/Transformation/w6
Contents
Transformation
pcpcG2 electro-transformation
In the evening of Aug. 10
Put electric tubes into 60℃ dryer for at least 2 hours, sterilized them by UV before electrotransformation.
Collect the cyanobacteria cell and diluted the cell with sterilized ddH2O washed it for 5 times,then diluted it with sterile ddH2O, mixed with pcpcg2-HR plasmid, transfered them into the electric tube, tunk for several times so that the liquid can reach to the bottom, installed the tube into switch, did electro-transformation, then transfer the mixture to BG11 culture
In the evening of Aug. 11
Centrifuged the cyanobacteria, diluted the cell with 400ul liquid culture, mixed with 5ml soft agar BG11 medium, spread on solid medium which contained 20 micrograms kanamycin per milliliter, put the plate into illumination incubator at 30 ℃.