Monday 27th 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 digested using HincII restriction enzyme.
| 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.
| 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 (DS-NMcas), SP dCas9 (DS-SPcasN-), ST1 dCas9 (DS-ST1casN-) and Td dCas9 (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.
