Difference between revisions of "Team:Paris Saclay/Notebook/July/5"

(→‎Transformation of DH5α|pPS16_004 and DH5α|pPS16_007)
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6 clones of each of the other transformations [[Team:Paris_Saclay/Notebook/June/28#pPS16_001|pPS16_001, pPS16_002, pPS16_003, pPS16_005 and pPS16_006]] were grown in 4mL of LB with Ampicillin (50µg/mL) at 37°C, 180rpm.  For pPS16_002 only 2 clones were cultivated because there was not more colony. To obtain more colonies, another [[Team:Paris_Saclay/Experiments#HeatShockCompetent|transformation]] was made with 50µL of cells and 5µL of plasmid. 50µL of transformed bacteria were plated on LB + Ampicillin (50µg/mL).
 
6 clones of each of the other transformations [[Team:Paris_Saclay/Notebook/June/28#pPS16_001|pPS16_001, pPS16_002, pPS16_003, pPS16_005 and pPS16_006]] were grown in 4mL of LB with Ampicillin (50µg/mL) at 37°C, 180rpm.  For pPS16_002 only 2 clones were cultivated because there was not more colony. To obtain more colonies, another [[Team:Paris_Saclay/Experiments#HeatShockCompetent|transformation]] was made with 50µL of cells and 5µL of plasmid. 50µL of transformed bacteria were plated on LB + Ampicillin (50µg/mL).
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===Biobrick characterization===
 
===Biobrick characterization===

Revision as of 14:32, 13 July 2016

Tuesday 5th July

Lab work

Visualization

Transformation of DH5α|pPS16_004 and DH5α|pPS16_007

By Caroline and Mathilde

The 04/06/2016 transformations show white colony growth for gBlocks 4.2 clone 3 and GFP1-9 (clone 1), but blue colonies only for 4.1 clone 3.

Thus this last gBlock 4.1 clone 3 was transformed again following the same protocol as the 21/06/2016 with : 50μg of competent DH5α cells 4.1 ligation product in 5μL of the plasmid puc19 4.1 clone 6 extracted DNA (which presented a good size strip on electrophoresis)

6 clones of each of the other transformations pPS16_001, pPS16_002, pPS16_003, pPS16_005 and pPS16_006 were grown in 4mL of LB with Ampicillin (50µg/mL) at 37°C, 180rpm. For pPS16_002 only 2 clones were cultivated because there was not more colony. To obtain more colonies, another transformation was made with 50µL of cells and 5µL of plasmid. 50µL of transformed bacteria were plated on LB + Ampicillin (50µg/mL).


Biobrick characterization

B-Galactosidase and luciferase test on transformed BL21

By Charlene

Cultures tested

Plasmid(s) K1372001 K1372001 + pcl UAA K1372001 + pcl UAG K1372001 + pcl Tq
Clone 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2
Salicylate concentration 0 30µM 1mM 0 30µM 1mM 0 30µM 1mM 0 30µM 1mM
Ratio between luciferase activity and β-Gal activity





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