Difference between revisions of "Team:Paris Saclay/Notebook/June/27"
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| − | =Monday | + | =Monday 27<sup>th</sup> June= |
| − | == | + | |
| − | ''' | + | ===Interlab study=== |
| − | + | ====Cell culture==== | |
| − | + | ''By Lea'' | |
| + | |||
| + | 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. | ||
| + | |||
| + | pUC19 plasmid was [[Team:Paris_Saclay/Experiments#PlasmidDigestion|digested]] using HincII restriction enzyme. | ||
| + | |||
| + | {| class="wikitable" | ||
| + | |- | ||
| + | ! Component | ||
| + | ! Volume (µL) | ||
| + | |- | ||
| + | | Plasmid | ||
| + | | 5 | ||
| + | |- | ||
| + | | Tango buffer 1x | ||
| + | | 5 | ||
| + | |- | ||
| + | | Water | ||
| + | | 38 | ||
| + | |- | ||
| + | | Enzyme | ||
| + | | 1 | ||
| + | |} | ||
| + | |||
| + | The mix was incubated for 1 hour at 37°C. A migration on electrophoresis gel (0.8% of agarose)was done for 30s. | ||
| + | |||
| + | {| class="wikitable" | ||
| + | |- | ||
| + | ! Component | ||
| + | ! Volume (µL) | ||
| + | |- | ||
| + | | Digested DNA | ||
| + | | 5 | ||
| + | |- | ||
| + | | Water | ||
| + | | 5 | ||
| + | |- | ||
| + | | Loading buffer | ||
| + | | 2 | ||
| + | |} | ||
| + | A DNA-ladder was used to visualize the DNA fragments size. | ||
| + | |||
| + | ===Bringing DNA closer=== | ||
| + | ====Cell culture==== | ||
| + | ''By Alice and Lea'' | ||
| + | |||
| + | 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. | ||
| + | |||
| + | 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. | ||
| + | |||
| + | ===BioBrick characterization=== | ||
| + | ''By Alice and Lea'' | ||
| + | |||
| + | BioBrick [http://parts.igem.org/Part:BBa_K1372001 K1372001] from iGEM team Paris Saclay 2014 was chosen to be characterized. | ||
| + | |||
| + | 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
