Template:SJTU-BioX-Shanghai/Notebook/Transformation/w5

Transformation

pdark natural transformation

In the noon of Aug. 5

Inoculated wildtype cyanobacteria into the flask and make sure the initial OD was about 0.1;

In the evening of Aug. 7

When the OD of our culture reached 0.4-0.6, collect the cell and mix the DNA, cyanobacteria and liquid medium, meanwhile set a negative control culture (without DNA), put them into illumination incubator at 30℃;

Aug. 7

Shook by hand for about 4 times from morning（every 2-3 hours）, after 8 hours, took 200ul culture of each tube, spread them on different solid plates which contained 20 micrograms kanamycin per milliliter, put the plate into illumination incubator at 30℃.

pcpcG2 electro-transformation

At the noon of Aug. 8

We inoculated wildtype cyanobacteria into the flask and make sure the initial OD was about 0.1; prepare the electric tubes