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

(BioBrick construction)
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{{Team:Paris_Saclay/notebook_header}}
 
{{Team:Paris_Saclay/notebook_header}}
=Monday 27th July=
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=Monday 27<sup>th</sup> June=
==Meeting==
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'''Members present:'''
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* Instructors and advisors: Claire, Fabio, Marie, Olivier, Sylvie, Philippe.
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* Students: Carla, Caroline, Charlène, Claire, Coline, Léa, Mahnaz, Marion, Martin, Maxence, Naiane, Yacine
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==Labwork==
 
 
===Interlab study===
 
===Interlab study===
 
====Cell culture====
 
====Cell culture====
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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.  
  
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===Visualization===
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====Visualization====
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''By Caroline, Charlène and Naïane''
  
===Visualisation===
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gBlocks from IDT were received on the 24/06/2016.
====BioBrick construction====
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''By Caroline, Charlène and Naïane"
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G-block from IDT were received on the 24/06/2016.
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pUC19 plasmid was [[Team:Paris_Saclay/Experiments#PlasmidDigestion|digested]] using HincII restriction enzyme.
 
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Puc19 plasmid was digested using HincII restriction enzyme.
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{| class="wikitable"
 
{| class="wikitable"
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|}
 
|}
  
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.
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The mix was incubated for 1 hour at 37°C. A migration on electrophoresis gel (0.8% of agarose)was done for 30s.
An electrophoresis was done (0.8% of agarose).
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{| class="wikitable"
 
{| class="wikitable"
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| 5
 
| 5
 
|-
 
|-
| Electrophoresis buffer
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| Loading buffer
 
| 2
 
| 2
 
|}
 
|}
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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 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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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====
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===BioBrick characterization===
 
''By Alice and Lea''
 
''By Alice and Lea''
  
BioBrick K1372001 from iGEM team Paris Saclay 2014 was chosen to be characterized.
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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

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.