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{{Team:Paris_Saclay/notebook_header}} | {{Team:Paris_Saclay/notebook_header}} | ||
=Wednesday 6<sup>th</sup> July= | =Wednesday 6<sup>th</sup> July= | ||
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====Preparation of LB solid and liquid stock==== | ====Preparation of LB solid and liquid stock==== | ||
''By Léa, Naiane, Laetitia'' | ''By Léa, Naiane, Laetitia'' | ||
− | - 2L of LB liquid : 20g/L powder LB + 2L water μQ | + | - 2L of LB liquid : 20g/L powder LB + 2L water μQ<br/> |
− | - 1L of LB solid : 1L of LB liquid +15g/L of Agar | + | - 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. |
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===Biobrick characterization=== | ===Biobrick characterization=== | ||
− | ==== ==== | + | ==== DNA extraction of K1372001 clone 1 and clone 2==== |
− | ''By '' | + | ''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 with the speedvac. The extractions were kept at -20°C. | ||
===Visualization=== | ===Visualization=== | ||
− | ==== | + | ==== gBlocks PCR screening ==== |
''By Caroline'' | ''By Caroline'' | ||
− | + | 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. | |
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− | + | 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 ==== | |
+ | ''By Léa, Caroline and Laetitia'' | ||
+ | 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