Notebook

You can find a short overview over our activities over this iGEM Season


March-May


Brainstorming and Literature Research ( Everyone): The entire team took the time from the first meeting to the beginning of lab work to define our project, do literature research, discuss ideas and solidify roles and responsibilities. We consulted our advisors and PIs every step of the way.

Fundraising and Contacting of sponsors ( Sandra, Tatyana, Zahra, Jakob): As we settled on project idea, we immediately brainstormed possible sponsors and interested parties. Under the leadership of Sandra, we began contacting sponsors, went to project pitches, collected feedback, and integrated it into our project design.

Communication and Collaborations ( Sandra, Magdalena, Neha, Paula, Tatyana): Parallel to all other activities, our team has began brainstorming a social media strategy, possible collaborations, and looked for iGEM teams to collaborate with on the ideas they propose.


June


Lab Safety Instructions ( Sandra, Magdalena, Julia, Tatyana, Anna, Irfan, Paula): The team member that were set to work in the wet lab recieves their safety instructions so they could work at the lab responsibly.

Fundraising and Contacting of sponsors ( Sandra, Tatyana, Zahra, Jakob): Our work on securing sponsorships continues.

Communication and Collaborations ( Sandra, Magdalena, Neha, Paula, Tatyana): Our first posts on social media are being developed. Template for the newsletter is made. Team logo and project logo is designed. First collaboration meetings commense.

Plasmid Design (Julia, Tatyana): After having settled on potetnial coding sequences, we began the work of codon-optimization, ordering parts and conceptualizing protocols for our lab work in July.


July


Lab Work Timeline

04.07.-10.07

Media Preparation and Pouring of Plates (Paula, Magdalena & Irfan): In the beginning, it was important to stock up on all the important things like media and agar that we could immediately use once we had our necessary strains.

Inventory (Paula, Julia & Anna): We had to take inventory of everything we had available before beginning work

11.07.-17.07.

Media Preparation and Pouring of Plates (Paula, Magdalena & Irfan): In the beginning, it was important to stock up on all the important things like media and agar that we could immediately use once we had our necessary strains.

Gel Pouring (Paula, Julia, Magdalena &Irfan): Every week, the wetlab team stocked up on clean-up and analytical gels.

Spider Silk: PCR Amplification (Paula, Julia): Part T, S#1, S#2 & S#3

Gel Electrophoresis, Sequencing (Paula, Julia & Magdalena): Analysis of results of PCRs

18.07-24.07.

Spider Silk: PCR Amplification (Anna, Julia & Magdalena): MaSp_A, MaSp_B, MaSp_C.

Media Preparation and Pouring of Plates (Anna, Magdalena & Irfan): Throughout the project, it was important to keep up continuous inventory.

Gel Pouring (Julia, Magdalena &Irfan): Every week, the wetlab team stocked up on clean-up and analytical gels.

Gel Electrophoresis, Sequencing (Anna, Julia & Magdalena): Analysis of results of PCRs

25.07.-31.07

Spider Silk: PCR Amplification (Julia): MaSp_A, MaSp_B, MaSp_C.

Media Preparation and Pouring of Plates (Anna, Magdalena & Irfan): Throughout the project, it was important to keep up continuous inventory.

Gel Pouring (Anna, Magdalena &Irfan): Every week, the wetlab team stocked up on clean-up and analytical gels.

Gel Electrophoresis, Sequencing (Julia & Anna): Analysis of results of PCRs

Other Activities in July

Collaborations: In Person Meet-up (Magdalena & Zahra): European iGEM meetup in Hamburg.

Partnerships (Tatyana, Magdalena and Sandra): First meetings of our various fruitful partnerships.

Communication (Neha & Magdalena): Continuous work on our Social Media.

August


01.08.-07.08

Amplifying Gelatin parts via PCR (Magdalena).

Golden Gate Cloning Backbone 1 (Julia): First Trials; MaSp insert + secretion factor

Transformation into E.coli (Julia): BB1 with MaSp insert

Media Preparation and Pouring of Plates (Tatyana, Magdalena & Irfan): Throughout the project, it was important to keep up continuous inventory.

Gel Pouring (Tatyana, Julia, Magdalena &Irfan): Every week, the wetlab team stocked up on clean-up and analytical gels.

Mini-Prep, Restriction Digest & Sequencing (Julia): MaSp insert backbone 1

08.08.-14.08

Amplifying Gelatin parts via PCR (Magdalena).

Golden Gate Cloning Backbone 1 (Julia): MaSp insert + secretion factor

Transformation into E.coli (Julia): BB1 MaSp insert

Mini-Prep, Restriction Digest & Sequencing (Julia): MaSp insert backbone 1

09.08: Brick Experiments (Sandra): 31 bricks created according to protocol; size 1x1x1

15.08.-21.08

Amplifying Gelatin parts via PCR (Magdalena)

Golden Gate Cloning Backbone 1 (Julia): MaSp insert + secretion factor

Transformation into E.coli (Julia): BB1 MaSp insert

Mini-Prep, Restriction Digest & Sequencing (Julia): MaSp insert backbone 1

Media Preparation and Pouring of Plates (Magdalena & Irfan): Throughout the project, it was important to keep up continuous inventory.

Gel Pouring (Tatyana & Irfan): Every week, the wetlab team stocked up on clean-up and analytical gels.

17.08.: Material Testing (Paula): According to protocol. Testing of bricks from 9.8. Results described here

Brick Experiments (Sandra & Paula) 106 Bricks created according to protocol; size 1x1x1 and 3.5x3.5x2.5.

22.08.-28.08.

Amplifying Gelatin parts via PCR (Magdalena).

Golden Gate Cloning: Backbone 2 and Backbone 3 (Julia): 4 variants of BB2 and 2 variants of BB3 for P.pastoris

Transformation into E.coli (Julia): BB2 & BB3 MaSp insert

Mini-Prep, Restriction Digest (Julia): MaSp insert BB2 & BB3

23.08.:Material Testing (Paula): According to protocol. Testing of bricks from 19.8. Results described here

Media Preparation and Pouring of Plates (Magdalena & Irfan): Throughout the project, it was important to keep up continuous inventory.

Gel Pouring (Tatyana & Irfan): Every week, the wetlab team stocked up on clean-up and analytical gels.

29.08.-31.08.

Trial: Fusing gelatin insert via PCR (Magdalena).

Golden Gate Cloning: Backbone 2 and Backbone 3 (Julia): 4 variants of BB2 and 2 variants of BB3 for P.pastoris

Transformation into E.coli (Julia): BB2 & BB3 MaSp insert

Mini-Prep, Restriction Digest (Julia): MaSp insert BB2 & BB3

30.08.: Bio-Glue (Sandra): Zoom conference at 14:00 with Prof. Gübitz for possible collaboration and advice on the use of lignin.

Other Activities in August

Fundraising (Sandra, Zahra & Tatyana): Project pitches, conversations with potential sponsors. Arrangement of future material testing.

Collaborations: In Person Meet-up (Magdalena, Paula & Neha): Junior Jam in Münster.

Collaborations: Podcast Appearance (Tatyana & Julia): iGEM Exeter "Test Tube" Podcast.

Partnerships (Tatyana, Magdalena and Sandra): Progress on ongoing partnerships.

Communication (Neha & Magdalena): Continuous work on our Social Media.

September


01.09.-04.09.

Trial: Fusing gelatin insert via PCR (Magdalena).

Golden Gate Cloning: Backbone 2 and Backbone 3 (Julia): 4 variants of BB2 and 2 variants of BB3 for P.pastoris

Transformation into E.coli (Julia): BB2 & BB3 MaSp insert

Mini-Prep, Restriction Digest (Julia): MaSp insert BB2 & BB3

Media Preparation and Pouring of Plates (Tatyana, Magdalena & Irfan): Throughout the project, it was important to keep up continuous inventory.

Gel Pouring (Tatyana & Irfan): Every week, the wetlab team stocked up on clean-up and analytical gels.

05.09.-11.09.

Trial: Fusing gelatin insert via PCR (Magdalena).

Golden Gate Backbone 3 (Julia): 4 expression casettes for MaSp P.pastoris

Mini-Prep, Restriction Digest (Julia): 4x MaSp BB3

Transformation into E.coli (Julia): 4x MaSp BB3

08.09.: Bio-Glue (Paula): Zoom conference at 14:00 with Ing. Sandra Bischof for possible collaboration and advice on the use of lignin.

Media Preparation and Pouring of Plates (Tatyana, Magdalena & Irfan): Throughout the project, it was important to keep up continuous inventory.

Gel Pouring (Tatyana & Irfan): Every week, the wetlab team stocked up on clean-up and analytical gels.

12.09.-18.09.

Trial: Fusing gelatin insert via Gibson Assembly (Magdalena).

Golden Gate Backbone 3 (Julia): 4 expression casettes for MaSp P.pastoris

Mini-Prep, Restriction Digest (Julia): 4x MaSp BB3

Golden Gate Backbone 1 (Julia): for PHB P. pastoris

Transformation into E.coli (Julia): PHB BB1

Mini-Prep, Restriction Digest, Sequencing (Julia): PHB insert BB1

Media Preparation and Pouring of Plates (Tatyana & Magdalena): Throughout the project, it was important to keep up continuous inventory.

Gel Pouring (Tatyana & Magdalena): Every week, the wetlab team stocked up on clean-up and analytical gels.

19.09.-25.09.

Trial: Fusing gelatin insert via NEBs Gibson Assembly (Magdalena).

Golden Gate Backbone 2 (Julia): for PHB in P. pastoris

Transformation of P. pastoris (Julia): 1xMaSp BB3, 2 variants

Transformation into E.coli (Julia): PHB in BB2

Mini-Prep, Restriction Digest (Julia): PHB insert BB2

25.09.: Brick Experiments (Sandra & Paula): 12 Bricks created according to protocol; size 3.5x3.5x2.5.

Media Preparation and Pouring of Plates (Tatyana, Zahra & Magdalena): Throughout the project, it was important to keep up continuous inventory.

Gel Pouring (Tatyana & Zahra): Every week, the wetlab team stocked up on clean-up and analytical gels.

26.09.-30.09.

Golden Gate Backbone 1 (Magdalena & Zahra): for gelatin in BB1.

Golden Gate Backbone 3 (Julia): for PHB in P. pastoris

Transformation into E.coli (Julia): PHB in BB3

Mini-Prep, Restriction Digest (Magdalena, Julia & Zahra): PHB insert BB3; Gelatin in BB1

Transformation of P. pastoris (Julia): 1xMaSp BB3, 2 variants; 4x MaSp; PHB

28.09.: Bio-Glue (Paula): Visit in the laboratory in BOKU Tulln; 6 bricks created with added BioGlue; 100ml each.

Media Preparation and Pouring of Plates (Tatyana, Magdalena & Irfan): Throughout the project, it was important to keep up continuous inventory.

Gel Pouring (Tatyana & Irfan): Every week, the wetlab team stocked up on clean-up and analytical gels.

Other Activities in September

Project Modeling (Neha & Jakob): Brainstorming, structuring, and making of our models.

Fundraising (Sandra, Zahra & Tatyana): Project pitches, conversations with potential sponsors. Arrangement of future material testing.

Collaborations: In Person Meet-up (Magdalena): Conference in Düsseldorf.

Collaborations: Podcast Appearance (Tatyana & Julia): iGEM Exeter "Test Tube" Podcast.

Partnerships (Tatyana & Magdalena): Progress on ongoing partnerships.

Communication (Neha, Tatyana & Magdalena): Continuous work on our Social Media.

Wiki Programming (Tatyana): Programming the layout of the wiki, writing Wiki Texts

Wiki Writing (Everyone): writing wiki Texts

October


01.10-02.10.

Golden Gate Backbone 1 (Magdalena): for gelatin in BB1.

Golden Gate Backbone 3 (Julia): for PHB in P. pastoris

Transformation into E.coli (Julia): PHB

Mini-Prep, Restriction Digest (Magdalena, Julia & Zahra): PHB insert BB3; gelatin in BB1

Transformation of P. pastoris (Julia): 1xMaSp BB3, 2 variants; 4x MaSp; PHB

Engineered P.pastoris cultivation (Julia): 1x MaSp insert, 2 variants

Media Preparation and Pouring of Plates (Tatyana, Zahra & Irfan): Throughout the project, it was important to keep up continuous inventory.

Gel Pouring (Tatyana & Zahra): Every week, the wetlab team stocked up on clean-up and analytical gels.

03.10.-09.10.

Golden Gate Backbone 3 (Magdalena & Zahra): for gelatin in BB3.

Transformation into E.coli (Magdalena): gelatin in BB3

Mini-Prep, Restriction Digest (Magdalena & Zahra): gelatin in BB3

Engineered P.pastoris cultivation (Julia): 1x MaSp insert, 2 variants; 4x MaSp insert; PHB

SDS-PAGE (Julia): supernatant of P.pastoris + 1x MaSp variant inserts culture.

10.10.Brick Creation and Material Testing (Paula): Mixture 4 brick creation; Brick testing.

Media Preparation and Pouring of Plates (Tatyana, Zahra & Irfan): Throughout the project, it was important to keep up continuous inventory.

Gel Pouring (Tatyana & Zahra): Every week, the wetlab team stocked up on clean-up and analytical gels.

Other Activities in September

Project Modeling (Neha & Jakob): Finishing the models, describing the results.

Partnerships (Tatyana & Magdalena): Last meetings of ongoing partnerships.

Communication (Neha, Tatyana & Magdalena): Continuous work on our Social Media.

Wiki Programming (Tatyana): Programming the layout of the wiki, writing wiki texts

Wiki Writing (Everyone): writing wiki texts