Week of July 18th
07.20 Wednesday
Transform acs and CreiLOV
Restriction digest on acs and CreiLOV
Meet with Dr. Chin
07.21 Thursday
Overnight cultures
07.22 Friday
Meet with Toulouse iGEM Team
Midi-prep on overnight cultures
Make glycerol stocks from overnight cultures
Week of July 25th
07.25 Monday
Midi-prep of acs and CreiLOV
07.26 Tuesday
Midi-prep on acs and CreiLOV
07.27 Wednesday
PCR of acs
Week of August 29th
09.02 Friday
Transform E. coli with IDT parts (iaaM, ami1, LPAAT, DGAT 1, DGAT 2, GPAT 1, GPAT 2) Plate
Plasmid = pUCIDT-AMP Genes = iaaM, ami1, LPAAT, DGAT1, DGAT2, GPAT1, GPAT2
09.03 Saturday
Run PCR reaction on colonies with IDT parts
Week of September 5th
09.05 Monday
Overnight cultures
Run gel with DNA from PCR from 09.03
09.06 Tuesday
Midi preps on overnight cultures
09.07 Wednesday
Restriction digest on GPAT 1 & 2 and DGAT 1 & 2
Restriction digest on iaaM and ami1 to put into pSB1C3
Restriction digest on promoter, RBS, and terminator from kit plates for expression in E coli
09.08 Thursday
PCR on midi prep from 09.06
Ligate GPAT 1 & 2 and DGAT 1 & 2
Ligate genes for IAA operon (order: promoter, RBS, iaaM, RBS, ami1, terminator)
09.09 Friday
Gel on midi prep PCR from 09.08
Debrief on experimental progress
09.10 Saturday
- digest parts 2022 kit plate 1
Promoter in PSB1C3SA plasmid
Promoter + GFP in PSB1C3 (could not find only the promoter in PSB1C3)
RBS in pSB1C3C
Terminator in pSB1C3SD
- Digest parts 2021 kit plate 1 (I think we should use these if we can find the kit plates)
Terminator in PSB1C3
Promoter in pSB1C3
RBS in pSB1C3
Week of September 12th
09.12 Monday
Couldn’t find gel stain :(
Run gel with DNA from PCR from 09.03
09.13 Tuesday
Transform promoter, RBS, and terminator parts into E. coli
Restriction digest on a plasmid containing GUS gene from Synbiodinium project
Ask Dr. Brewer about steps to Western Blot
Look for tryptophan, Western Blot equipment, and competent cell stock in the lab
09.14 Wednesday
Overnight culture on promoter, RBS, terminator
Make LB Broth
The Gel stain has arrived in the lab :)
Week of September 12th
09.12 Monday
Couldn’t find gel stain :(
Run gel with DNA from PCR from 09.03
09.13 Tuesday
Transform promoter, RBS, and terminator parts into E. coli
Restriction digest on a plasmid containing GUS gene from Synbiodinium project
Ask Dr. Brewer about steps to Western Blot
Look for tryptophan, Western Blot equipment, and competent cell stock in the lab
09.14 Wednesday
Overnight culture on promoter, RBS, terminator
Make LB Broth
The Gel stain has arrived in the lab :)
09.15 Thursday
MIDI (not mini, should have 500 ml LB) prep on the promoter, RBS, terminator
Protocol
09.16 Friday
Restriction digest on promoter, RBS, terminator
Ligate IAA sequence (order: promoter, RBS, iaaM, RBS, ami1, terminator)
09.17 Saturday
Transform IAA sequence into E. coli (using rosetta cells for expression, not the usual electrocompetent cells)
Ask Eric what data is needed for the IAA model
09.18 Sunday
Overnight culture on 1A 1, 1i 1,15E 1, 13M 2
Gel electrophoresis and gel extraction on DGAT 1 & 2 and GPAT 1 & 2
Week of September 19th
Monday 09.19
Midi prep on 1A 1, 1i 1,15E 1, 13M 2
Overnight culture on 1A 2, 15E 2, 16O 1, 16O 2, 13M 1
Wednesday 09.21
Restriction digest on 1A and 1i
Thursday 09.22
Midi prep on ___________.
Friday 09.23
Midi prep on 1A1 and 1I1
Restriction digest on plasmids from 09.22
Ligate promoter part into plasmids
Week of September 19th
Monday 09.26
Midi prep on iaaM and ami1
Overnight culture on pBI121
Tuesday 09.27
Record concentrations of results on midi prep on iaaM and ami1
No midi prep needed for pBI121 (algae plasmid)
Restriction digest on iaaM and ami1
Ligate iaaM and ami1 with promoter
Meeting with Dr. Knoshaug
Wednesday 09.28
Transform complete iaaM and ami1 construct into E. coli
Check lab equipment for Western blot (transfer machine and shaker)
Thursday 09.29
Overnight culture on iaaM and ami1 constructs
Colony PCR on plates from 09.28
Friday 09.30
Start Western blot (no western blot done today)
Saturday 10.01
Finish Western blot
