Difference between revisions of "Team:Paris Saclay/Notebook/September/21"
(Created page with "{{Team:Paris_Saclay/notebook_header}} = Wednesday 21<sup>st</sup> September= ==Lab work== ===Visualization=== ====Colony PCR of 16 clones containing GFP 1.9 in pSB1C3 (pPS16...") |
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= Wednesday 21<sup>st</sup> September= | = Wednesday 21<sup>st</sup> September= | ||
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===Visualization=== | ===Visualization=== | ||
| − | ====Colony PCR of 16 clones containing GFP | + | ====Colony PCR of 16 clones containing corrected dCas9 ST - GFP 11 in pSB1C3 (pPS16_017)==== |
| − | ''By Maxence | + | ''By Maxence, Mahnaz & Coline'' |
A colony PCR was done for 16 clones from the 20th September. For that purpose, 16 clones were screened and used for the usual [[Team:Paris_Saclay/Experiments##Polymerase_chain_reaction|protocol]] of Colony PCR. The same colonies were also plated on Petri dish containing solid LB + 30 µg/mL Cm and liquid cultures (LB + 30 µg/mL Cm) were made from these clones. Both petri dishes and liquid cultures were grown at 37°C. | A colony PCR was done for 16 clones from the 20th September. For that purpose, 16 clones were screened and used for the usual [[Team:Paris_Saclay/Experiments##Polymerase_chain_reaction|protocol]] of Colony PCR. The same colonies were also plated on Petri dish containing solid LB + 30 µg/mL Cm and liquid cultures (LB + 30 µg/mL Cm) were made from these clones. Both petri dishes and liquid cultures were grown at 37°C. | ||
| Line 27: | Line 27: | ||
|3 min | |3 min | ||
|- | |- | ||
| − | |rowspan="3"| | + | |rowspan="3"|25 cycles |
|95°C | |95°C | ||
|30 sec | |30 sec | ||
|- | |- | ||
| − | | | + | |48.4°C |
|30 sec | |30 sec | ||
|- | |- | ||
|72°C | |72°C | ||
| − | | | + | |4min |
|- | |- | ||
|Final Extension | |Final Extension | ||
| Line 46: | Line 46: | ||
|} | |} | ||
| − | + | [[Team:Paris_Saclay/Experiments#primers|Primers]] used were: | |
{| class="wikitable" | {| class="wikitable" | ||
|- | |- | ||
!Matrix | !Matrix | ||
| − | !Clones containing GFP | + | !Clones containing corrected dCas9 ST - GFP 11 in pSB1C3 (pPS16_017) |
|- | |- | ||
|Primers | |Primers | ||
| − | | | + | |iPS168 and iPS169 |
|- | |- | ||
|} | |} | ||
| Line 67: | Line 67: | ||
!Expected band size (bp) | !Expected band size (bp) | ||
|- | |- | ||
| − | |GFP | + | |dCas9 ST - GFP 11 |
| − | | | + | |3800 |
|} | |} | ||
| + | [[File:T--Paris Saclay--Gel111111.png|400px|thumb|center|Result of the migration]] | ||
{{Team:Paris_Saclay/notebook_footer}} | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 17:09, 9 October 2016
