(→B-Galactosidase and luciferase test on transformed BL21)
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* 2µL of loading dye
* 2µL of loading dye
* 5 µL of the digestion product
* 5 µL of the digestion product
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* 5 µL of H2O
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* 5 µL of water
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[[File:T--Paris_Saclay--160705_bringingDNAcloser_casdigestion_échelle.jpg|400px|thumb|right|DS-NMcas, DS-SPcasN-, DS-ST1casN- and DS-TDcasN- digestion (Migration)]]
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[[File:T--Paris_Saclay--160705_bringingDNAcloser_casdigestion_échelle1.JPG|400px|thumb|right|DS-NMcas, DS-SPcasN-, DS-ST1casN- and DS-TDcasN- digestion (Migration)]]
===Biobrick characterization===
===Biobrick characterization===
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|colspan="6"|K1372001
|colspan="6"|K1372001
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|colspan="6"|K1372001 + pcl UAA
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|colspan="6"|K1372001 + pcl_TAA
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|colspan="6"|K1372001 + pcl UAG
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|colspan="6"|K1372001 + pcl_TAG
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|colspan="6"|K1372001 + pcl Tq
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|colspan="6"|K1372001 + pcl_Tq
|-
|-
!Clone
!Clone
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|}
|}
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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.
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''By Alice and Laetitia''
''By Alice and Laetitia''
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10 µL of K1372001 two clones culture was put in culture in 3mL of LB + 3µL of chloramphenicol. Cultures were incubated at 37°c, 180 rpm.
+
10 µL of cultures of the two clones expressing K1372001 were put in culture in 3mL of LB + 3µL of chloramphenicol. Cultures were incubated at 37°C, 180 rpm.
