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| | ==Lab work== | | ==Lab work== |
| | ===Visualization=== | | ===Visualization=== |
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| − | ==== Glycerol stocks of clones X, X, X and X containing corrected dCas9 ST - GFP 11 in pSB1C3 (pPS16_017)====
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| − | "By Maxence, Mahnaz & Coline"
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| − | The glycerol stock of the bacteria with the following plasmids were made.
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| − | *pPS16_017 (dCas9 ST - GFP 11) clone X
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| − | *pPS16_017 (dCas9 ST - GFP 11) clone X
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| − | *pPS16_017 (dCas9 ST - GFP 11) clone X
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| − | *pPS16_017 (dCas9 ST - GFP 11) clone X
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| − | 500µL of liquid culture and 250 µL of glycerol 60% were mixed and put at -20°C.
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| − | ====Plasmids extraction of clones X, X, X and X containing corrected dCas9 ST - GFP 11 in pSB1C3 (pPS16_017)====
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| − | "By Maxence, Mahnaz & Coline"
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| − | The following plasmids were extracted using the [[Team:Paris_Saclay/Experiments#InvitrogenPlasmidExtraction|"Charge Switch-Pro Plasmid Miniprep"]] kit:
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| − | *pPS16_017 (dCas9 ST - GFP 11) clone X
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| − | *pPS16_017 (dCas9 ST - GFP 11) clone X
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| − | *pPS16_017 (dCas9 ST - GFP 11) clone X
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| − | *pPS16_017 (dCas9 ST - GFP 11) clone X
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| − |
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| − | ====Digestion of plasmids extracted from clones X, X, X and X containing corrected dCas9 ST - GFP 11 in pSB1C3 (pPS16_017)====
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| − | "By Maxence, Mahnaz & Coline"
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| − | To assess if DH5a cells were transformed with the good plasmid, plasmid digestion was run with XbaI as following:
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| − | * 1 µL of extracted plasmid
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| − | * 1 µL of buffer
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| − | * 1 µL of restriction enzyme XbaI
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| − | * 7 µL of water
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| − | A control was done with 1 µL of non-digested plasmid.
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| − | GEL
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| | ====Colony PCR of 16 clones containing FRB - GFP 11 - GFP 1.9 in pSB1C3 (pPS16_022)==== | | ====Colony PCR of 16 clones containing FRB - GFP 11 - GFP 1.9 in pSB1C3 (pPS16_022)==== |
