Difference between revisions of "Team:Paris Saclay/Notebook/June/27"
(→BioBrick construction) |
|||
| (20 intermediate revisions by 6 users not shown) | |||
| Line 1: | Line 1: | ||
{{Team:Paris_Saclay/notebook_header}} | {{Team:Paris_Saclay/notebook_header}} | ||
| − | =Monday | + | =Monday 27<sup>th</sup> June= |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
| − | |||
| − | |||
===Interlab study=== | ===Interlab study=== | ||
====Cell culture==== | ====Cell culture==== | ||
| Line 14: | Line 8: | ||
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. | |
| − | + | ||
| − | + | ||
| − | + | pUC19 plasmid was [[Team:Paris_Saclay/Experiments#PlasmidDigestion|digested]] using HincII restriction enzyme. | |
| − | + | ||
| − | + | ||
{| class="wikitable" | {| class="wikitable" | ||
| Line 41: | Line 34: | ||
|} | |} | ||
| − | 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. |
| − | + | ||
{| class="wikitable" | {| class="wikitable" | ||
| Line 55: | Line 47: | ||
| 5 | | 5 | ||
|- | |- | ||
| − | | | + | | Loading buffer |
| 2 | | 2 | ||
|} | |} | ||
| Line 64: | Line 56: | ||
''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. |
| + | |||
| + | 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. |
| + | |||
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
