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

(Created page with "{{Team:Paris_Saclay/notebook_header}} =Monday 27th July= ==Meeting== '''Members present:''' * Instructors and advisors: Claire, Fabio, Marie, Olivier, Sylvie, Philippe. * Stud...")
 
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* Instructors and advisors: Claire, Fabio, Marie, Olivier, Sylvie, Philippe.
 
* Instructors and advisors: Claire, Fabio, Marie, Olivier, Sylvie, Philippe.
 
* Students: Carla, Caroline, Charlène, Claire, Coline, Léa, Mahnaz, Marion, Martin, Maxence, Naiane, Yacine
 
* 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.
 +
 +
 +
===Visualisation===
 +
====BioBrick construction====
 +
''By Caroline, Charlène and Naïane"
 +
 +
G-block from IDT were received on the 24/06/2016.
 +
 +
Puc19 plasmid was digested using HincII restriction enzyme.
 +
 +
{| class="wikitable"
 +
|-
 +
! Component
 +
! Volume (µL)
 +
|-
 +
| Plasmid
 +
| Tango buffer 1x
 +
| Water
 +
| Enzyme
 +
|-
 +
| 5
 +
| 5
 +
| 38
 +
| 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).
 +
 +
{| class="wikitable"
 +
|-
 +
! Component
 +
! Volume (µL)
 +
|-
 +
| Digested DNA
 +
| Water
 +
| Electrophoresis buffer
 +
|-
 +
| 5
 +
| 5
 +
| 2
 +
|}
 +
A DNA-ladder was used.
 +
 +
===Bringing DNA closer===
 +
====Cell culture====
 +
''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.
 +
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.
  
 
{{Team:Paris_Saclay/notebook_footer}}
 
{{Team:Paris_Saclay/notebook_footer}}

Revision as of 13:36, 4 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.


Visualisation

BioBrick construction

By Caroline, Charlène and Naïane"

G-block from IDT were received on the 24/06/2016.

Puc19 plasmid was digested using HincII restriction enzyme.

Component Volume (µL)
Plasmid Tango buffer 1x Water Enzyme
5 5 38 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 Water Electrophoresis buffer
5 5 2

A DNA-ladder was used.

Bringing DNA closer

Cell culture

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. 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.





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