Difference between revisions of "Template:SJTU-BioX-Shanghai/Notebook/Transformation/w5"
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====pdark natural 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; | 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℃; | 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''' | + | '''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℃. | 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℃. | ||
Revision as of 08:27, 16 September 2015
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 8th, Aug
We inoculated wildtype cyanobacteria into the flask and make sure the initial OD was about 0.1; prepare the electric tubes