Difference between revisions of "Team:Paris Saclay/Notebook/June/21"
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{{Team:Paris_Saclay/notebook_header}} | {{Team:Paris_Saclay/notebook_header}} | ||
=Tuesday 21<sup>st</sup> June= | =Tuesday 21<sup>st</sup> June= | ||
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| − | ==== | + | ====Heat-shock competent cells preparation==== |
| − | '' | + | ''By Lea, Caroline and Marion'' |
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| + | Culture OD was measured at 600nm (OD<sub>600nm</sub> = 3.4). | ||
| + | 3.4mL were put into 250mL of LB medium and incubated at 20°C and 180rpm overnight. | ||
===Interlab study=== | ===Interlab study=== | ||
| − | ====Transformation==== | + | ====Transformation (cf. [[Team:Paris_Saclay/Experiments#HeatShockCompetent|protocole]])==== |
| − | '' | + | ''By Lea, Caroline and Marion'' |
50µL of competent DH5α cells were transformed with 1µL of plasmids (for each construction: device 1, 2, 3, negative control and positive control) and one control tube was made without plasmids. Tubes were kept into ice for 30min then a thermal shock was made at 42°C for 1min. 500µL LB medium was added into each tube and incubated at 37°C for one 1h. | 50µL of competent DH5α cells were transformed with 1µL of plasmids (for each construction: device 1, 2, 3, negative control and positive control) and one control tube was made without plasmids. Tubes were kept into ice for 30min then a thermal shock was made at 42°C for 1min. 500µL LB medium was added into each tube and incubated at 37°C for one 1h. | ||
| − | Petri | + | Petri dishes were prepared with selective medium containing LB, agar and 30µg/mL chloramphenicol. |
| − | Each transformation condition was | + | Each transformation condition was spread on Petri dishes in duplicate and incubated overnight at 37°C. |
{{Team:Paris_Saclay/notebook_footer}} | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 13:04, 9 October 2016
