Difference between revisions of "Team:Paris Saclay/Notebook/August/18"

(→‎Thursday 18th August)
(→‎B-Galactosidase and luciferase test on transformed BL21)
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''By Charlene and Mathilde''
 
''By Charlene and Mathilde''
  
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Cultures were spun down for 5min at 1300rpm. Supernatant was discarded and glass marbles previously cleaned with 1M nitric acid were added with 50μL of Luc buffer. Tubes were vortexed for 30min at 4°C. Everything was then done on ice. 150μL of Luc bufer was added and cells were centrifuged for 15min at 13000rom at 4°C.
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Cultures were spun down for 5min at 13000rpm. Supernatant was discarded and glass marbles previously cleaned with 1M nitric acid were added with 50μL of Luc buffer. Tubes were vortexed for 30min at 4°C. Everything was then done on ice. 150μL of Luc bufer was added and cells were centrifuged for 15min at 13000rpm at 4°C.
  
 
For the luciferase test, 4mL of Luc buffer were mixed to 80μL of ATP and 8μL of luciferine.
 
For the luciferase test, 4mL of Luc buffer were mixed to 80μL of ATP and 8μL of luciferine.
 
For the βGal test, 400μL of Z buffer mixed with 10μL of extract and 100μL of ONPG were incubated for 8min at 30°C. Then 250μL of STOP buffer was added.
 
For the βGal test, 400μL of Z buffer mixed with 10μL of extract and 100μL of ONPG were incubated for 8min at 30°C. Then 250μL of STOP buffer was added.
 
OD<sub>420nm</sub> was measured.
 
OD<sub>420nm</sub> was measured.

Revision as of 12:16, 18 August 2016

Thursday 18th August

Lab work

Visualization

gBlocks 3 digestion with Eco47III

By Alice

Biobrick characterization

B-Galactosidase and luciferase test on transformed BL21

By Charlene and Mathilde

Cultures were spun down for 5min at 13000rpm. Supernatant was discarded and glass marbles previously cleaned with 1M nitric acid were added with 50μL of Luc buffer. Tubes were vortexed for 30min at 4°C. Everything was then done on ice. 150μL of Luc bufer was added and cells were centrifuged for 15min at 13000rpm at 4°C.

For the luciferase test, 4mL of Luc buffer were mixed to 80μL of ATP and 8μL of luciferine. For the βGal test, 400μL of Z buffer mixed with 10μL of extract and 100μL of ONPG were incubated for 8min at 30°C. Then 250μL of STOP buffer was added. OD420nm was measured.