Difference between revisions of "Team:Paris Saclay/Notebook/July/29"
(Created page with "{{Team:Paris_Saclay/notebook_header}} = Thursday 28<sup>th</sup> July= ==Lab work== ===Visualization=== ==== ==== ''By Caroline '' ==== ==== ''By '' {{Team:Paris_Saclay/no...") |
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{{Team:Paris_Saclay/notebook_header}} | {{Team:Paris_Saclay/notebook_header}} | ||
| − | = | + | = Friday 29<sup>th</sup> July= |
| − | + | ||
===Visualization=== | ===Visualization=== | ||
| − | ==== ==== | + | ====gBlock 1.1, 1.2 and GFP1-9 insertion in puc19==== |
| − | ''By Caroline '' | + | ''By Caroline'' |
| + | |||
| + | The insertion was carried out following the usual [[Team:Paris_Saclay/Experiments#Ligation|protocol]]. | ||
| + | |||
| + | ====High fidelity PCR on bacteria transformed with [[Team:Paris_Saclay/Notebook/June/28#pPS16_001|pPS16_001]], [[Team:Paris_Saclay/Notebook/June/28#pPS16_002|pPS16_002]] and [[Team:Paris_Saclay/Notebook/June/28#pPS16_005|pPS16_005]] ==== | ||
| + | ''By Alice and Mathilde '' | ||
| + | |||
| + | A PCR with Q5® High-Fidelity 2X Master Mix was performed on clones selected after screening PCR made on July 28, following [[Team:Paris_Saclay/Experiments#Q5PCR|this protocol]]. We choose clones which had the good size of insert. We selected the clone 2 for DH5α|pPS16_001; clones 4,7 and 8 for DH5α|pPS16_002 and clones 1 and 4 DH5α|pPS16_005. [[Team:Paris_Saclay/Experiments#primers|1151_pheoR and 1152_pheoF]] primers were used. Annealing temperature was 66°C. After amplification, 1 µL of loading dye was added to 5µL of each PCR products. Then 5 µL of this mix were put on gel. PCR products were migrated 30min at 100V. | ||
| − | + | PCR products expected were : | |
| − | + | ||
| + | {| class="wikitable" | ||
| + | |- | ||
| + | !Plasmids | ||
| + | !Band size (bp) | ||
| + | |- | ||
| + | |pPS16_001 | ||
| + | |1017 | ||
| + | |- | ||
| + | |pPS16_002 | ||
| + | |1017 | ||
| + | |- | ||
| + | |pPS16_005 | ||
| + | |1017 | ||
| + | |} | ||
| + | [[File:T--Paris Saclay--160729 Visualisation PCR3.jpeg|400px|thumb|right|Migration of gBlocks PCR product]] | ||
{{Team:Paris_Saclay/notebook_footer}} | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 16:13, 9 October 2016
