Team:Paris Saclay/Notebook/August/28
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(Created page with "{{Team:Paris_Saclay/incl_debut_generique}} ='''Notebook : August 27'''= =='''summary'''== *We got clonies on ligation promoter fnr(activator)nirB plus RBS_LacZ+Term_PSB1C3 ...")
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(Created page with "{{Team:Paris_Saclay/incl_debut_generique}} ='''Notebook : August 27'''= =='''summary'''== *We got clonies on ligation promoter fnr(activator)nirB plus RBS_LacZ+Term_PSB1C3 ...")
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Revision as of 15:21, 12 September 2013
Notebook : August 27
summary
- We got clonies on ligation promoter fnr(activator)nirB plus RBS_LacZ+Term_PSB1C3
promoter fnr(activator)nirB plus RBS_AmilCP+Term_PSB1C3 promoter fnr(repressor) plus RBS_LacZ+Term_PSB1C3 promoter fnr(repressor) plus RBS_AmilCP+Term_PSB1C3t so we do Colony PCR for it
- We didn't get clonies on Gibson so we do a gel electrophoresis of parts of Gibson assembly to verify the size
- Digestion Dnp1 purification of the the PSB1C3 clean for Gibson and electrophoresis of it .
lab work
A - Aerobic/Anaerobic regulation system / B - PCB sensing system
- 1 - Gel extraction of the PSB1C3 Clean for Gibson.
Protocol : Gel extraction
- 2- Migration of 3µl PSB1C3 Clean for Gibson (product gel extraction) '
[[]] |
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- 3- product quantification.
- We used the nano drop to measure the DNA in 260nm and we found its concentration
name | PSB1C3 Clean dig Dnp1 gel extraction |
conc. | 37.2ng/µl |
- 4 - Gibson assembly.
- RBS_BphR2_part1, BphR2_part2 and plasmid PSB1C3
- FNR_part1, FNR part1 and plasmid PSB1C3
- RBS_FNR part1, FNR_part2 and plasmid PSB1C3
Sample Volume:
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These 3 mixture is incubated at 50°C for up to one hour. Transformation and spread in 6 LB plates with Chlorenphenicol . Incubate at 37°C over night.