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

(Lab Meeting)
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''By Alice and Lea''
 
''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.  
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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.  
 
Plasmids were delivered into tubes containing LB agar medium and bacteria colony containing plasmids.  

Revision as of 08:20, 16 October 2016

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 ([1]), SP dCas9 ([2]), ST1 dCas9 ([3]) and Td dCas9 ([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 [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.