Difference between revisions of "Team:Paris Saclay/Notebook/June/21"

(Interlab study)
(Lab work)
 
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{{Team:Paris_Saclay/notebook_header}}
 
=Tuesday 21<sup>st</sup> June=
 
=Tuesday 21<sup>st</sup> June=
==Lab work==
 
  
====Competent cells preparation====
+
====Heat-shock competent cells preparation====
 
''By Lea, Caroline and Marion''
 
''By Lea, Caroline and Marion''
  
Culture OD was measured at 600nm (OD = 3.4).  
+
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.  
+
3.4mL were put into 250mL of LB medium and incubated at 20°C and 180rpm overnight.
 
+
  
 
===Interlab study===
 
===Interlab study===
Line 15: Line 13:
  
 
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 dish were prepared with selective medium containing LB, agar and 30µg/mL chloramphenicol.  
+
Petri dishes were prepared with selective medium containing LB, agar and 30µg/mL chloramphenicol.  
Each transformation condition was displayed into petri dish in duplicate and then incubated at 37°C overnight.  
+
Each transformation condition was spread on Petri dishes in duplicate and incubated overnight at 37°C.  
  
 
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{{Team:Paris_Saclay/notebook_footer}}

Latest revision as of 13:04, 9 October 2016

Tuesday 21st June

Heat-shock competent cells preparation

By Lea, Caroline and Marion

Culture OD was measured at 600nm (OD600nm = 3.4). 3.4mL were put into 250mL of LB medium and incubated at 20°C and 180rpm overnight.

Interlab study

Transformation (cf. 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. Petri dishes were prepared with selective medium containing LB, agar and 30µg/mL chloramphenicol. Each transformation condition was spread on Petri dishes in duplicate and incubated overnight at 37°C.