Difference between revisions of "Team:Paris Saclay/Notebook/June/27"
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{{Team:Paris_Saclay/notebook_header}} | {{Team:Paris_Saclay/notebook_header}} | ||
| − | =Monday | + | =Monday 27<sup>th</sup> June= |
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===Interlab study=== | ===Interlab study=== | ||
====Cell culture==== | ====Cell culture==== | ||
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Transformed bacteria frozen on 24/06/2016 were put into 3mL of LB medium containing 30µg/mL chloramphenicol and incubated overnight at 37°C, 180rpm. | Transformed bacteria frozen on 24/06/2016 were put into 3mL of LB medium containing 30µg/mL chloramphenicol and incubated overnight at 37°C, 180rpm. | ||
| + | ===Visualization=== | ||
| + | ====Visualization==== | ||
| + | ''By Caroline, Charlène and Naïane'' | ||
| − | + | gBlocks from IDT were received on the 24/06/2016. | |
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| − | + | pUC19 plasmid was [[Team:Paris_Saclay/Experiments#PlasmidDigestion|digested]] using HincII restriction enzyme. | |
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| − | The mix was incubated for 1 hour at 37°C. | + | The mix was incubated for 1 hour at 37°C. A migration on electrophoresis gel (0.8% of agarose)was done for 30s. |
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| − | | | + | | Loading buffer |
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| − | A DNA-ladder was used. | + | A DNA-ladder was used to visualize the DNA fragments size. |
===Bringing DNA closer=== | ===Bringing DNA closer=== | ||
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''By Alice and Lea'' | ''By Alice and Lea'' | ||
| − | 4 plasmids containing NM Cas9, SP | + | 4 plasmids containing NM Cas9 ([https://www.addgene.org/48646/ DS-NMcas]), SP dCas9 ([https://www.addgene.org/48657/ DS-SPcasN-]), ST1 dCas9 ([https://www.addgene.org/48659/ DS-ST1casN-]) and Td dCas9 ([https://www.addgene.org/48660/ DS-TDcasN-]) were ordered to Addgene and received on 24/06/2016. |
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| + | Plasmids were delivered into tubes containing LB agar medium and bacteria colony containing plasmids. | ||
Bacteria were put into 15mL of LB medium containing 50µg/mL of spectinomycin and incubated overnight at 37°C, 180 rpm. | Bacteria were put into 15mL of LB medium containing 50µg/mL of spectinomycin and incubated overnight at 37°C, 180 rpm. | ||
| − | + | ===BioBrick characterization=== | |
''By Alice and Lea'' | ''By Alice and Lea'' | ||
| − | BioBrick K1372001 from iGEM team Paris Saclay 2014 was chosen to be characterized. | + | BioBrick [http://parts.igem.org/Part:BBa_K1372001 K1372001] from iGEM team Paris Saclay 2014 was chosen to be characterized. |
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10µL of DH5α|K1372001 cells were put into 3mL of LB medium containing 30µg/mL of chloramphenicol and incubated overnight at 37°C, 180 rpm. | 10µL of DH5α|K1372001 cells were put into 3mL of LB medium containing 30µg/mL of chloramphenicol and incubated overnight at 37°C, 180 rpm. | ||
{{Team:Paris_Saclay/notebook_footer}} | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 08:20, 16 October 2016
