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

Line 5: Line 5:
 
===Visualization===
 
===Visualization===
 
====Transformation of DH5α|pPS16_004 and DH5α|pPS16_007 ====
 
====Transformation of DH5α|pPS16_004 and DH5α|pPS16_007 ====
−
 
+
''By Caroline and Mathilde''
  
 
The 04/06/2016 transformations show white colony growth for G-blocks 4.2 clone 3 and GFP1-9  (clone 1), but blue colonies only for 4.1 clone 3.
 
The 04/06/2016 transformations show white colony growth for G-blocks 4.2 clone 3 and GFP1-9  (clone 1), but blue colonies only for 4.1 clone 3.
Line 14: Line 14:
 
4.1 clone 6 extracted DNA (which presented a good size strip on electrophoresis)
 
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 placed in 4mL of LB added to Ampicillin (50µg/mL) at 37°C, 180RPM.  For 2.2 just 2 clones were cultivated because there was not more colony.  
+
6 clones of each of the other transformations pPS16_001, pPS16_002, pPS16_003, pPS16_005 and pPS16_006 were placed in 4mL of LB added to Ampicillin (50µg/mL) at 37°C, 180RPM.  For 2.2 just 2 clones were cultivated because there was not more colony. To obtain more colonies, another transformation (Heat shock competent cells, section protocols) was made with 50µL of cells, 5µL of plasmid. 50µL of transformed bacteria were displayed on LB + Ampicillin (50µg/mL)dishes.
 +
 
 +
===Biobrick characterization===
 +
====B-Galactosidase and luciferase test on transformed BL21====
 +
''By Charlene''
 +
 
 +
'''Cultures tested'''
 +
{| class="wikitable"
 +
|-
 +
|Plasmid(s)
 +
|K1372001
 +
|K1372001
 +
|K1372001
 +
|K1372001
 +
|K1372001
 +
|K1372001
 +
|K1372001 + pcl UAA
 +
|K1372001 + pcl UAA
 +
|K1372001 + pcl UAA
 +
|K1372001 + pcl UAA
 +
|K1372001 + pcl UAA
 +
|K1372001 + pcl UAA
 +
|K1372001 + pcl UAG
 +
|K1372001 + pcl UAG
 +
|K1372001 + pcl UAG
 +
|K1372001 + pcl UAG
 +
|K1372001 + pcl UAG
 +
|K1372001 + pcl UAG
 +
|K1372001 + pcl Tq
 +
|K1372001 + pcl Tq
 +
|K1372001 + pcl Tq
 +
|K1372001 + pcl Tq
 +
|K1372001 + pcl Tq
 +
|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
 +
|0
 +
|30µM
 +
|30µM
 +
|1mM
 +
|1mM
 +
|0
 +
|0
 +
|30µM
 +
|30µM
 +
|1mM
 +
|1mM
 +
|0
 +
|0
 +
|30µM
 +
|30µM
 +
|1mM
 +
|1mM
 +
|}
 
   
 
   
 
{{Team:Paris_Saclay/notebook_footer}}
 
{{Team:Paris_Saclay/notebook_footer}}

Revision as of 13:59, 12 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 G-blocks 4.2 clone 3 and GFP1-9 (clone 1), but blue colonies only for 4.1 clone 3.

Thus this last G-block 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 placed in 4mL of LB added to Ampicillin (50µg/mL) at 37°C, 180RPM. For 2.2 just 2 clones were cultivated because there was not more colony. To obtain more colonies, another transformation (Heat shock competent cells, section protocols) was made with 50µL of cells, 5µL of plasmid. 50µL of transformed bacteria were displayed on LB + Ampicillin (50µg/mL)dishes.

Biobrick characterization

B-Galactosidase and luciferase test on transformed BL21

By Charlene

Cultures tested

Plasmid(s) K1372001 K1372001 K1372001 K1372001 K1372001 K1372001 K1372001 + pcl UAA K1372001 + pcl UAA K1372001 + pcl UAA K1372001 + pcl UAA K1372001 + pcl UAA K1372001 + pcl UAA K1372001 + pcl UAG K1372001 + pcl UAG K1372001 + pcl UAG K1372001 + pcl UAG K1372001 + pcl UAG K1372001 + pcl UAG K1372001 + pcl Tq K1372001 + pcl Tq K1372001 + pcl Tq K1372001 + pcl Tq K1372001 + pcl Tq 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 0 30µM 30µM 1mM 1mM 0 0 30µM 30µM 1mM 1mM 0 0 30µM 30µM 1mM 1mM





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