Difference between revisions of "Team:Paris Saclay/Notebook/June/27"

(Labwork)
(Cell culture)
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''By Alice and Lea''
 
''By Alice and Lea''
  
4 plasmids containing NM Cas9, SP Cas9, ST1 Cas9 and Td Cas9 were ordered to addgene and received on the 24/06/2016. Plasmids were delivered into tubes containing LB agar medium and bacteria colony containing plasmids.  
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4 plasmids containing NM Cas9 ([https://www.addgene.org/48646/|DS-NMcas]), SP Cas9 ([https://www.addgene.org/48657/|DS-SPcasN-]), ST1 Cas9 ([https://www.addgene.org/48659/|DS-ST1casN-]) and Td Cas9 ([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===
 
===BioBrick characterization===

Revision as of 09:08, 12 July 2016

Monday 27th July

Meeting

Members present:

  • Instructors and advisors: Claire, Fabio, Marie, Olivier, Sylvie, Philippe.
  • Students: Carla, Caroline, Charlène, Claire, Coline, Léa, Mahnaz, Marion, Martin, Maxence, Naiane, Yacine


Labwork

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

BioBrick construction

By Caroline, Charlène and Naïane

GBlock 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. Then, 1µL of HincII was added and the mix was incubated for 1 hour at 37°C. An electrophoresis was done (0.8% of agarose).

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 ([1]), SP Cas9 ([2]), ST1 Cas9 ([3]) and Td Cas9 ([4]) 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 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.