Team:NAWI Graz/Notebook
October
- Team assembly with guidance from the previous team
- creation of a facebook site and exchange of contact data
- setting up of a weekly meet up appointment
first introduction to the competition and team assembly
second meet up with members of the 2017 team; contact data exchange; creation of various communication platforms and setting up of a weekly meet-up appointment;
November
- start of brainstorming sessions to find the project we wanted to realize
- first deliberations about funding of the project
- first contacts about lab and office space
December
- team members with already assigned tasks left the team because of university obligations or personal reasons, sizing us down to 6 people from previously 18
- still ongoing project finding
January
- asking people in the respective fields about the viability of our collected project ideas
- deciding on the palm kernel oil project since it is such a huge discussion point in austria
February
- creation of a budget plan (Anna, Marlene)
- presentation of our project idea to the deanery of the universities TU-Graz and KFU-Graz
March
- search for a lab space for the preliminary tests (Thomas)
- extensive change to the project design (esterification)
- gain of 3 new members raising our number to 9
Movie night with following presentation of our project at the university (Anna, Marlene, Marco, Henrik, Veronika, Thomas)
April
- improved budget plan
- planing of the new preliminary tests and laboratory allocation (Marco, Dominik, Thomas)
- igem-graz.at was created and set (Jakob)
lange nacht der forschung - information evening for visitors of the universities or research facilities
Teambuilding evening - the team got together for a game of escape the room
May
- preliminary yeast tests
- first two weeks: WT and elo knock out strain
- rest of the month: checking the extracts of other strains for better compatibility.(Marco, Dominik, Thomas)
first day of the preliminary tests
Mol-Bio Fest'l - annually public event at the institut to gain more attention for our project and the iGEM competition and celebrate the achievements
June
- Labwork came to a halt as a result of other obligations at the university since this is a busy time (labcourses and exams)
- contacting a lot of companies for possible cooperations (Anna, Marlene, Veronika, Birgit)
Teambuilding evening
TU-Graz StreeTech - public outreach event in front of the city hall Graz to present all student competition teams and projects of the TU Graz including the iGEM teams NAWI_Graz 2017 and 2018 (Anna, Marlene, Henrik, Veronika, Thomas)
BHAK Weiz - public outreach in form of a presentation and small DNA extraction experiments in a public school to gain more attention for our field of study and iGEM itself (Anna, Marlene, Veronika)
July
- creation of the polycistronic expression cassette in snapgene, to be implemented with CRISPR-Cas9 and lambda red recombineering into E. coli K12 MG1655 at the position of the dgkA gene – knocking it out in the process. The creation took some time since the correct use of snapgene had to be learned and several meetings with professionals from ACIB where needed. They explained to us the correct way to create an expression casette for our chosen system (two-vector system proposed by Jiang et.al. In 2015 ) and the most common errors that could occur. (Thomas, Dominik)
Project presentation at Biomin - a local animal nutirion company (Marlene, Birgit, Thomas)
iGEM meetup munich
iGEM meetup munich
iGEM meetup munich
August/September/October
- Main Labwork (Marco, Dominik, Thomas)
- Programming/Design of the Wiki Page (Jakob)
OE-PCR of gblocks 1+2, 3+4, 5+6, 7+8, 9+10 with limited success
OE-PCR of only the first 2 gblocks under different conditions to improve the yield of combined gblocks.
OE-PCR reaction of the first set of gblocks with a diffrent protocol that was suggested by professionals at ACIB; agarose gel electrophoresis to check the results of our PCR;
innoculation of YPD medium with delta-desaturase pre-culture; cutting the gblock bands from our agarose gel and storing them in freezer;
yeast batch was not grown enough and kept in the shaker
yeast batch was still at a low cell count and incubated for another day
determination of the cell count and microscopy of the delta desaturase strain to check on the condition of the cells; harvest of the cells and freezing them with liquid nitrogen for storage in a -80°C freezer till lipidextraction is in order.
plating of the plasmid carrying bacteria on agarose plates for the creation of a glycerol stock for long term storage.
restricition digest of plasmid backbone psB1K3 and the gblocks tesA mutated C12/C14, atfA old and atfA optimized;
inactivation of restriction enzymes; ligation of the gblocks into the psB1K3 backbone
gel purification of the stored agarose bands; desalting of the ligation mix; colony PCR for amplification of the weak bands from the gel;
OE-PCR tesA, gblock1-2, gblock3-4, gblock9-10
agarose gel purification of tesA and 1+2, 3+4; nanodrop measurement
OE-pcr gblocks 1+2, 3+4, 7+8 ;
1+2+3+4 oe-pcr and gelpurification
Interlab Marburg begin
preparation for PCR and two vector system
cPCR
Agarose gel electrophoresis, gel purification
PCR (different thermocycler), electrophoresis, purification
Nanodrop, PCR (better thermocycler again), electrophoresis, purification Nanodrop
restriction digest, ligation, preparation of spec and spec/kan media ONC of plasmid stocks
miniprep, double restriction digest, ligation, gel elctrophoresis, single digest
gel electrophoresis, double digest (longer time)?
tranformation, double restriction digest, ligation, miniprep, Yale conference
cultivation of electrocompetent cells, transforamation, miniprep, ligation
ONCs, gel elctrophoresis
minipreps, gel electrophoresis, gel purification, PCR
PCR, restriction digest, ligation, gel elctrophoresis
PCR, gel-electrophoresis, transformation, ONCs, TU-welcome days - a public outreach event for graduates to find their future studys
single colony streaks, ONCs, (successful biobrick implementation), TU-welcome days
miniprep; getting biobricks ready for transport
Spritzerstand'l - a typical public celebration at our university for especially students and our form to prepare for the final project part, the "horror"-wiki week
“Dirndl und Lederhosen” photoshoot - a local seller of traditional clothing helped us to produce our final team pictures and clearly represent our (educationally) homeland Styria as a part of Austria
ongoing work on the wiki page
ongoing wotk on the wiki page
first events for the new team assembly
finalisation of the wiki page
heading of to vienna and checking in for our flight to Boston - see you all there!
end of log
