Difference between revisions of "Team:Paris Saclay/Notebook/October/6"
(→BioBrick X characterization) |
(→Lab work) |
||
| (4 intermediate revisions by one other user not shown) | |||
| Line 2: | Line 2: | ||
= Thursday 6<sup>th</sup> October= | = Thursday 6<sup>th</sup> October= | ||
| − | + | ||
===Visualization=== | ===Visualization=== | ||
| Line 14: | Line 14: | ||
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. | 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. | |
====Transformation of DH5a cells with and FKBP - GFP 10 (pPS16_018) and FRB - GFP 11 (pPS16_019)==== | ====Transformation of DH5a cells with and FKBP - GFP 10 (pPS16_018) and FRB - GFP 11 (pPS16_019)==== | ||
| Line 21: | Line 21: | ||
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]]. | 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=== | |
| − | ===BioBrick | + | |
====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==== | ====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'' | ''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 | + | 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_018 (FKBP - GFP 10) clone 6 | ||
Latest revision as of 15:55, 14 October 2016
