Difference between revisions of "Team:Paris Saclay/Notebook/June/30"

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The mix was incubated for 1 hour at 37°C and then stored at -20°C over the night.
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The mix was incubated for 1 hour at 37°C and then stored at -20°C until the gel electrophoresis was done. Fragments are expected to be as follow:
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{| class="wikitable"
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|-
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!Plasmid name
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!Plasmid size (kb)
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!Digestion product size (kb)
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|-
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|1.1
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|3.7
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|2.7 and 1
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|-
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|1.2
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|3.7
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|2.7 and 1
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|-
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|2.1
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|3.7
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|2.7 and 1
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|-
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|3.1
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|3.7
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|2.7 and 1
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|-
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|3.2
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|3.7
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|2.7 and 1
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|-
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|4.1
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|3.4
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|2.7 and 0.7
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|-
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|4.2
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|4
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|2.7 and 1.3
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|-
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|GFP 1-9
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|3.6
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|2.7 and 0.9
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|}
  
 
===Biobrick characterization===
 
===Biobrick characterization===

Revision as of 16:37, 6 July 2016

Thursday 30th June

Lab work

Visualization

Extraction of the plasmids containing gBlocks

By Alice and Lea

DNA from bacteria transformed with plasmids containing gBlocks was extracted following the extraction protocol. For each gblock, DNA of 6 different clones was extracted. At the end, plamids were resuspended in water/RNAse (5µL RNAse 10 mg/mL for 1 mL of water) to send to sequencing.

Digestion of the plasmids containing gBlocks

By Alice and Léa

After extraction, plasmids were digested with EcoRI and HindIII.

Component Volume (µL)
Plasmids 3
Red buffer 10x 2
Water 14
EcoRI enzyme 0.5
HindIII enzyme 0.5

The mix was incubated for 1 hour at 37°C and then stored at -20°C until the gel electrophoresis was done. Fragments are expected to be as follow:

Plasmid name Plasmid size (kb) Digestion product size (kb)
1.1 3.7 2.7 and 1
1.2 3.7 2.7 and 1
2.1 3.7 2.7 and 1
3.1 3.7 2.7 and 1
3.2 3.7 2.7 and 1
4.1 3.4 2.7 and 0.7
4.2 4 2.7 and 1.3
GFP 1-9 3.6 2.7 and 0.9

Biobrick characterization

Electrocompetents and transformation by electroporation of BL21

By Caroline and Charlene

The same protocol as 29/06/16 was used for the plasmids K1372001, K1372001+pcl_TAA and the controls which were done following the same protocol but without plasmids. K1372001 and the controls were streaked on LB + Cm (30µg/mL). K137200 + PCLTAA was streaked on LB + Cm (30µg/mL) + Streptomicin (50µg/mL). This time two different streaks were made for each plasmid. The first one using 50µL from the culture. The second one was done using 100µL of each remaining culture (400µL) after concentration by centrifugation.

The petri dishes made on 29/06 were stored at 4ºC






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