Difference between revisions of "Team:Paris Saclay/Notebook/October/10"
(→Protein electrophoresis 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) |
(→Digestion of extracted plasmids by EcoRI) |
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= Monday 10<sup>th</sup> October= | = Monday 10<sup>th</sup> October= | ||
| − | == | + | |
| + | ===Visualization=== | ||
| + | |||
| + | ====Plasmids extraction 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 BbaB0015==== | ||
| + | ''By Sylvie'' | ||
| + | |||
| + | The OD were measured for each overnight cultures: | ||
| + | |||
| + | {| class="wikitable center" | ||
| + | |- | ||
| + | !Sample | ||
| + | !OD (1 ml) | ||
| + | |- | ||
| + | |FRB - GFP 11 in pSB1C3 (pPS16_019) | ||
| + | |3 | ||
| + | |- | ||
| + | |FKBP - GFP 10 in pSB1C3 (pPS16_018) | ||
| + | |3.9 | ||
| + | |- | ||
| + | |GFP 1.9 in pUC19 (pPS16_009) | ||
| + | |4.2 | ||
| + | |- | ||
| + | |BbaB0015 | ||
| + | |5 | ||
| + | |- | ||
| + | |} | ||
| + | |||
| + | |||
| + | Then the plasmids were extracted using a [[Team:Paris_Saclay/Experiments#Plasmid_DNA_extraction|"standard Plasmid Miniprep"]]. | ||
| + | |||
| + | ====Digestion of extracted plasmids by EcoRI==== | ||
| + | ''By Maxence & Caroline'' | ||
| + | |||
| + | Extracted pPS16_019, pPS16_018, pPS16_009 and BbaB0015 from the 10th October were digested by restriction enzymes EcoRI in order to verify the concentration as following: | ||
| + | |||
| + | * 2 µL of plasmid | ||
| + | * 2 µL of buffer FD | ||
| + | * 2 µL of restriction enzyme EcoRI | ||
| + | * 14 µL of water | ||
| + | |||
| + | The mix were incubated for 1 hour at 37°C. Then, 20 µL of digestion products and 15 µL of DNA ladder were placed in wells and migrated at 100V during 30 min. | ||
| + | |||
| + | [[File:T--Paris Saclay--Gelsylvie1.png|400px|thumb|center|Result of the migration]] | ||
| + | |||
| + | The digestions were good for pPS16_019, pPS16_018 and BbaB0015, but we did not obtained the expecting band size for pPS16_009. | ||
| + | |||
===BioBrick K2039000 and K2039001 characterization=== | ===BioBrick K2039000 and K2039001 characterization=== | ||
| Line 39: | Line 84: | ||
A protein electrophoresis of the extracted protein was done using the [[Team:Paris_Saclay/Experiments#Protein_electrophoresis_using_the_ThermoFisher_Mini_Gel_Tank.C2.AE_device|protein electrophoresis protocol]]. For that purpose, the gel cassette Bolt 4-12% Bis-Tris Plus 10W was used and the power was set for 1 hour at 65V. | A protein electrophoresis of the extracted protein was done using the [[Team:Paris_Saclay/Experiments#Protein_electrophoresis_using_the_ThermoFisher_Mini_Gel_Tank.C2.AE_device|protein electrophoresis protocol]]. For that purpose, the gel cassette Bolt 4-12% Bis-Tris Plus 10W was used and the power was set for 1 hour at 65V. | ||
| − | ==== | + | ====Transfert of protein extracted from 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'' | ''By Maxence'' | ||
Latest revision as of 14:51, 18 October 2016
