Difference between revisions of "Team:Paris Saclay/Notebook/October/6"
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= Thursday 6<sup>th</sup> October= | = Thursday 6<sup>th</sup> October= | ||
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===Visualization=== | ===Visualization=== | ||
| − | ==== | + | ====Plasmids extraction of FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4 and FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6==== |
| − | ''By Maxence'' | + | ''By Maxence & Victor'' |
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| + | The following plasmids were extracted using the [[Team:Paris_Saclay/Experiments#InvitrogenPlasmidExtraction|"Charge Switch-Pro Plasmid Miniprep"]] kit: | ||
| + | *pPS16_018 (FKBP - GFP 10) clone 6 | ||
| + | *pPS16_019 (FRB - GFP 11) clone 4 | ||
| − | + | After extraction, 5 µL of each plasmids and 20 µL of DNA ladder were placed in wells and migrated at 100V during 30 min in order to assess their concentration. | |
| − | + | No DNA has been extracted. | |
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| − | + | ====Transformation of DH5a cells with and FKBP - GFP 10 (pPS16_018) and FRB - GFP 11 (pPS16_019)==== | |
| − | + | ''By Maxence & Victor'' | |
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| − | [[Team:Paris_Saclay/Experiments# | + | As we were not able to obtain good plasmid concentration after culture of glycerol stocks and plasmid extraction, transformations were done. Dh5a cells were transformed with pSB1C3 containing FKBP - GFP 10 (pPS16_018) and FRB - GFP (pPS16_019) using the usual [[Team:Paris_Saclay/Experiments#heat-shocktransformation|protocol]]. |
| − | + | ===BioBrick K2039000 and K2039001 characterization=== | |
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| − | + | ====Culture of FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4, FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6, GFP 1.9 in pUC19 (pPS16_009) clone 1 and PhB1040 strain 594==== | |
| + | ''By Maxence & Victor'' | ||
| − | + | As we have met several issues in order to obtain plasmis pPS16_023 (gBlock ATG linker FRB + GFP 11 + gBlock ATG linker FKBP + GFP 10 + gBlock GFP 1-9), we decided to characterize our BioBricks with a Western-Blot with antibodies against GFP. For that purpose, the following clones were put on solid cultures (LB + 30 µg/mL Cm ou Amp) grown at 37°C overnight: | |
| − | + | *pPS16_018 (FKBP - GFP 10) clone 6 | |
| − | + | *pPS16_019 (FRB - GFP 11) clone 4 | |
| − | + | *pPS16_009 (GFP 1.9) clone 1 | |
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| − | + | Furthermore, another strain containing the full GFP was cultures in the same conditions: PhB1040 strain 594 (lac-3350 galk2 galT22 2psL179 pGFP) (Amp resistance). | |
{{Team:Paris_Saclay/notebook_footer}} | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 15:55, 14 October 2016
