Difference between revisions of "Team:Paris Saclay/Notebook/July/11"

(→‎DH5α transformation with DS-NMcas, DS-SPcasN-, DS-ST1casN- and DS-TDcasN-)
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Transformed cells were plated in duplicata (10μL and 50μL) on 8 Petri dishes (spectinomycin 50μg/mL). Control cells were plated on another plate (spectinomycin 50μg/mL).
 
Transformed cells were plated in duplicata (10μL and 50μL) on 8 Petri dishes (spectinomycin 50μg/mL). Control cells were plated on another plate (spectinomycin 50μg/mL).
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===Visualization===
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The same PCR as [[Team:Paris_Saclay/Notebook/July/8#Colony screening PCR|the 08/07/16]] was performed

Revision as of 15:34, 12 July 2016

Monday 11th July

Lab work

Biobrick characterization

Electrocompetents and transformation by electroporation of BL21

By Charlène

BL21 cells were grown up to OD600nm=7.14. They were then made competent and transformed with K1372001 extracted from clone 2 and pcl_TAA, pcl_TAG or pcl_Tq to create the following strains:

  • BL21|K1372001+pcl_TAA
  • BL21|K1372001+pcl_TAG
  • BL21|K1372001+pcl_Tq


Bringing DNA closer

DH5α transformation with DS-NMcas, DS-SPcasN-, DS-ST1casN- and DS-TDcasN-

By Laetitia and Mathilde

DS-NMcas, DS-SPcasN-, DS-ST1casN- and DS-TDcasN- plasmids were resuspended with 5μL of sterilized water. 5 tubes were prepared :

  • one control tube with only 50μL of DH5α heat-shock competent cells
  • four tubes with 50μL of DH5α heat-shock competent cells + 1μL of plasmid

The regular heat-shock competent cells transformation protocol was used.

Transformed cells were plated in duplicata (10μL and 50μL) on 8 Petri dishes (spectinomycin 50μg/mL). Control cells were plated on another plate (spectinomycin 50μg/mL).

Visualization

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The same PCR as the 08/07/16 was performed