Difference between revisions of "Team:Paris Saclay/Notebook/September/20"
(Created page with "{{Team:Paris_Saclay/notebook_header}} = Tuesday 20<sup>th</sup> September= ==Lab work== ===Visualization=== ====Samples preparation for sequencing==== "By Maxence, Mahnaz &...") |
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|FKBP - GFP 10 clone 10<div id="FKBP - GFP 10 clone 10"></div> | |FKBP - GFP 10 clone 10<div id="FKBP - GFP 10 clone 10"></div> | ||
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| + | |PCR product 1 obtained by iPS174 & iPS175<div id="PCR product 1 obtained by iPS174 & iPS175"></div> | ||
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| + | |PCR product 2 obtained by iPS173 & iPS176<div id="PCR product 2 obtained by iPS173 & iPS176"></div> | ||
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| + | ====Gibson of cleaned up PCR products from pSB1C3 with dCas9 ST - GFP 11 (pPS16_017) clone 8 treated by DpnI==== | ||
| + | ''By Maxence, Mahnaz, Coline & Caroline'' | ||
| + | |||
| + | Gibson was performed with cleaned up PCR products from pSB1C3 with dCas9 ST - GFP 11 (pPS16_017) clone 8 treated by DpnI, more precisely PCR product 1 obtained by iPS174 & iPS175 (plasmid) and PCR product 2 obtained by iPS173 & iPS176 (insert) with the following protocol: | ||
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| + | For each 20μl of reaction, mix the following reagents : | ||
| + | * 0.2 µL of insert | ||
| + | * 3.16 µL of plasmid | ||
| + | * 6.64 µL of water | ||
| + | * 10 µL of buffer mix | ||
| + | |||
| + | Two controls were done: one without insert and one without buffer (water was added in order to have a total of 20 µL). The PCR was performed as follow : 1 hour at 50°C. | ||
{{Team:Paris_Saclay/notebook_footer}} | {{Team:Paris_Saclay/notebook_footer}} | ||
Revision as of 15:50, 19 September 2016
