Difference between revisions of "Team:Paris Saclay/Notebook/July/6"
(→pPS16_004, pPS16_007 and the pPS16_007 clone 6 (selected the 1/07/2016) streaks) |
(→Lab work) |
||
| Line 1: | Line 1: | ||
{{Team:Paris_Saclay/notebook_header}} | {{Team:Paris_Saclay/notebook_header}} | ||
=Wednesday 6<sup>th</sup> July= | =Wednesday 6<sup>th</sup> July= | ||
| − | + | ||
| − | + | ||
====Preparation of LB solid and liquid stock==== | ====Preparation of LB solid and liquid stock==== | ||
''By Léa, Naiane, Laetitia'' | ''By Léa, Naiane, Laetitia'' | ||
| Line 8: | Line 7: | ||
- 2L of LB liquid : 20g/L powder LB + 2L water μQ<br/> | - 2L of LB liquid : 20g/L powder LB + 2L water μQ<br/> | ||
- 1L of LB solid : 1L of LB liquid +15g/L of Agar<br/> | - 1L of LB solid : 1L of LB liquid +15g/L of Agar<br/> | ||
| − | The | + | The solutions were put in autoclave for sterilization with the help of Sylvain. |
===Biobrick characterization=== | ===Biobrick characterization=== | ||
| Line 14: | Line 13: | ||
''By Mathilde'' | ''By Mathilde'' | ||
| − | Plasmids DNA were extracted following the [[Team:Paris_Saclay/Experiments#PlasmidExtraction|usual]] protocol except that isopropanol was used instead of 100% ethanol and that pellets were dried on the lab bench top instead of | + | Plasmids DNA were extracted following the [[Team:Paris_Saclay/Experiments#PlasmidExtraction|usual]] protocol except that isopropanol was used instead of 100% ethanol and that pellets were dried on the lab bench top instead of with the speedvac. The extractions were kept at -20°C. |
===Visualization=== | ===Visualization=== | ||
| Line 20: | Line 19: | ||
''By Caroline'' | ''By Caroline'' | ||
| − | A PCR screening was | + | A PCR screening was carried out on colonies of bacteria transformed with the gBlocks (6 clones for each except pPS16_004 for which it was only 2 clones). We use the Taq [[Team:Paris_Saclay/Experiments#taqPCR|usual]] protocol. We used puc19 [[Team:Paris_Saclay/Experiments#primers|primers]] 1151_pheoR and 1152_pheoF. |
| − | 1 μL of each culture were added for each | + | 1 μL of each culture were added for each gblock pPS16_001, pPS16_002, pPS16_003, pPS16_004, pPS16_005, pPS16_006 and pPS16_007. |
==== pPS16_004, pPS16_007 and the pPS16_007 clone 6 (selected on 01/07/2016) streaks ==== | ==== pPS16_004, pPS16_007 and the pPS16_007 clone 6 (selected on 01/07/2016) streaks ==== | ||
''By Léa, Caroline and Laetitia'' | ''By Léa, Caroline and Laetitia'' | ||
| − | The transformations carried out the 30/06/2016 were streaked again on LB Petri dishes containing 50µg/ml of | + | The transformations carried out the 30/06/2016 were streaked again on LB Petri dishes containing 50µg/ml of amplicilin on which X-Gal/IPTG was spread on (white/blue screen). The Petri dishes were incubated ON at 37°C. |
{{Team:Paris_Saclay/notebook_footer}} | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 14:12, 9 October 2016
