iGEM 2021 — Team Bielefeld-CeBiTec

[![](https://static.igem.org/mediawiki/2021/8/8b/HQ_page_logo.jpg)](https://2021.igem.org)

# Team:Bielefeld-CeBiTec/Notebook

[![Home](https://static.igem.org/mediawiki/2021/4/4b/T--Bielefeld-CeBiTec--21_PLANT_logo.png)](https://2021.igem.org/Team%3ABielefeld-CeBiTec)

[ ]

People

[Team](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Team)

[Attribution](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Attributions)

[Partners](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Sponsoring)

[Medal Criteria](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Medal_Criteria)

[Bielefeld](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Bielefeld)

Human Practices

[Overview](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Human_Practices_Overview)

[Human Practices](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Human_Practices)

[Education](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Education)

[Experts](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Experts)

[Communication](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Communication)

[Collaborations](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Collaborations)

iGEM Phototrophs

[Partnership](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Partnership)

[Contribution](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Contribution)

Lab

[Experiments](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Experiments)

[Parts](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Parts)

[Basic Part](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Basic_Part)

[Improve-a-Part](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Improve)

[Safety](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Safety)

[Notebook](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Notebook)

Results

[Overview](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Results_Overview)

[Hydroculture](https://2021.igem.org/Team%3ABielefeld-CeBiTec/results_hydroculture)

[Bacteria](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Bacteria)

[Tobacco](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Tobacco)

[Reporter](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Reporter)

Project

[Overview](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Overview)

[Description](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Description)

[Model](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Model)

[Proof of Concept](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Proof_Of_Concept)

[Implementation](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Implementation)

[Engineering](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Engineering)

[Chemicals](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Chemicals)

[Outlook](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Outlook)

# Notebook

# June

14.06. – 20.06.

**Getting started**

- This week we can finally get started in the lab and spend the week organising
the lab, preparing buffers, medium

- Chemocompetent E.coli DH5α cells and Heat shock transformation to test them
(Paul)

21.06. – 27.06.

**Gene syntheses arrive**

- The gene Synthesis for our signaling cascade in bacteria and tobacco arrived, so
we started our attempts to amplificate them (Louisa)

- Testing the competent cells through heat shock transformation (Paul, Lennart)

-Creating glycerin-stocks

28.06. – 04.07.

**Starting reporters**

-Sequencing of 35S:RUBY (Alignment fits perfectly)

-We started working on our reporter systems by sequencing our 35S:RUBY
plasmid, creating a glykostock and started to amplificate parts of the plasmid
(Paul)

-Isolating P16∆:XVE and 35S:RUBY (Louisa)
-Attempt to amplify RUBY in two fragments (Louisa)

# July

05.07. – 11.07.

**Isolation and amplification**

-Attempting to amplify fragments for tobacco and bacterial cascade (Louisa, Julia,
Marie)

-Isolation of pK7WG2 (Lucas)

-Amplification of lexA (Marie)

-Attempting to amplify pK7WG2 in two parts (Louisa)

12.07. – 18.07.

**Testing chemicals in bacteria**

Preliminary measurement of lethal concentrations on TDG, DIMP, DEMP, MPA
(Jacob)

-Testing the lethal concentrations of DEMP, MPA, DIMP, TDG on E.coli DH5a
expressing mRFP, through growth measurement in the biolector (Tim, Marvin,
Matthes)

-Further attempts to amplify fragments of the signaling cascade (Louisa)

19.07. – 25.07.

**Agrobacterium**

-We received agrobacterium and prepared medium
Attempting to amplify fragments of signaling cascade and optimise PCR
conditions (Louisa)

-Generating new chemocompetent cells with different protocol and testing them
with heat shock transformation (Louisa)

-Isolating SUPERR:sXVE (Paul)

-Attempting to join together the amplified RUBY amplificates through GeneSOEing
(Julia)

# August

02.08. – 08.08.

**Improve a part**

- We started our labwork for improve a part by cultivating DH5a containing
pRSETb with mCRISPRed and Isolating the plasmid for further cloning steps
(Lennart, Tim)

- starting cloning for in vitro tests and improve a part
Transformation of Agrobacterium with 35S:RUBY and lexA:RUBY

09.08. – 15.08.

**Agroinfiltration**

- agroinfiltration of N.benthamiana with 35S:RUBY and are quite happy
with the results

- Further cloning efforts for our signaling cascade, improve a
part (Tim, Louisa, Lucas, Marie)

- amplification of fragments for Mobius assembly (Paul)

16.08. – 22.08.

**Betalain-extraction from Tobacco**

- Betalain-extraction from tobacco plants infiltrated with RUBY (Paul,
Julia)

- Preparation of second hydroculture experiment (Matthes, Jacob)

- Start of cloning efforts for our library (Lennart)

- Further optimization attempts for amplification of our
signaling cascade and cloning efforts (Louisa)

23.08. – 29.08.

**First hydroculture**

- Addition of our chemicals to the first hydroculture with a longer
runtime and sampling and documentation in regular intervals (Jacob, Matthes)

-Gibson assembly of tobacco signaling casacade and screening efforts (Julia,
Louisa)

30.08. – 05.09.

**Mobius Assembly**

-Plasmid isolation and PCRs for mobius assembly (Julia, Marie)

-cloning efforts for improve a part (Tim, Lucas)
- screening and inducing clones from bacterial signaling cascade (Tim, Lucas)

# September

06.09. – 12.09.

**Expression of mCRISPRed**
-preparation of second hydroculture and start of freeze drying of the samples
from first culture (Matthes, Jacob)

- Amplification, and cloning of RBP, BTCA-R and DIMP-R for receptor expression
and library (Lennart, Tim)

-Expression and Purification of mCRISPRed (Lucas)

- Testing lethal BTCA-Concentrations for bacteria with the Biolector (Marvin)

13.09. – 19.09

**RNA-iso and GC-MS preparation**

- Agroinfiltration of Tobacco with 35S:RUBY, 35S:RUBY and pK7WG2-P4H2-GFP
(Julia, Marie, Paul)

- RNA-Isolation, Betalain-extraction and PAM-Measurment of infiltrated Tobacco
leaves and addition of estrogen

- Transformation of BL21 DE3 with DIMP-R and BTCA-R (Jonas, Lennart)

- Preparation of GC-MS samples in the lyophylle and cell homogenizer

20.09. – 26.09.

**GC/MS is running**
- Amplification of signaling cascade was optimised with touchdown-PCR (Tim)

- RNA-Isolation of infiltrated plants (Mareike)

- Amplification and Mobius-assembly of reporter constructs (Marie)

- GC/MS was set up and started (Jacob, Matthes)

- Expression and Purification of mRuby3 (Lucas)

27.09. – 03.10.

**Concluding improve a part**

- Bradford assay for mRuby3 and mCRISPRed and measuring of
fluorescence intensity at different pH conditions at the TECAN reader (Lucas)

– cultivating and expression of BTCA-R in BL21 DE3

- Agroinfiltration of tobacco with 35S:RUBY, 35S:RUBY + pk7WG2:P4H4
and negative control (Julia)

- Mobius Assembly of the tobacco signaling cascade
(Julia, Paul, Marie)

-Repeating lethal BTCA concentration tests in biolector (Marvin)

- Gibson assembly of bacterial signaling cascade and
screening with colony PCR (Tim)

- Induction of positive colonies with Arabinose and Ribose and
fluorescence measurement with TECAN (Tim)

- Preparation of infiltrated plant samples for metabolome analysis
(Marvin)

# October

04.10. – 10.10.

**Measuring fluorescence with CLSM**

– Metabolome extraction from infiltrated plant samples and
measurement in GC-MS (Marvin)

- RNA-isolation from infiltrated plant samples
Bacterial signaling cascade – chemotaxis tests with ribose, measuring
fluorescence of different of bacteria under different conditions (Tim)

– Expression of DIMP and purification of DIMP and BTCA under
native and deanatured conditions and running SDS-PAGE with samples from
purification (Lucas)

11.10. – 17.10.

**Testing BTCA-Receptor with SPR**

– testing of BTCA-R with surface plasmon resonance spectrometer
(Lucas,Tim)

– co-transformation of BL21 with pJOE:RBP & cascade plasmid
and induction of liquid cultures and repeating chemoxtaxis experiment (Tim)

– transformation of Agrobacterium with reporter and signaling
cascade constructs, hydroculture of tobacco with BTCA, infiltrating hydrocultured
plants with Agrobacterium carrying lexA:RUBY, ANTHOS and signaling cascade

18.10. – 21.10.

 **Characterising the signaling cascade**

– inducing co-transformed bacteria in liquid culture and
measuring fluorescence with superresolution microscopy (Tim)
– measuring and comparing growth of bacteria with mRuby3 and
mCRISPRed (Lucas)

---

[![Promega](https://static.igem.org/mediawiki/2021/1/14/T--Bielefeld-CeBiTec--promega-logo.png)](https://www.promega.de/)
[![Merck](https://static.igem.org/mediawiki/2021/1/17/T--Bielefeld-CeBiTec--merck-logo.png)](https://www.merckgroup.com/de)
[![Plasmid-Factory](https://static.igem.org/mediawiki/2021/d/dd/T--Bielefeld-CeBiTec--pf-logo.jpg)](https://plasmidfactory.com/)
[![SnapGene](https://static.igem.org/mediawiki/2021/d/d0/T--Bielefeld-CeBiTec--snapgene-logo.png)](https://www.snapgene.com/)
[![Sparkasse-Bielefeld](https://static.igem.org/mediawiki/2021/5/58/T--Bielefeld-CeBiTec--spkb-logo.png)](https://www.sparkasse-bielefeld.de/de/home.html)
[![Starlab](https://static.igem.org/mediawiki/2021/e/ec/T--Bielefeld-CeBiTec--starlab_logo.svg)](https://www.starlabgroup.com/)
[![Zymo](https://static.igem.org/mediawiki/2021/2/27/T--Bielefeld-CeBiTec--zymo-logo.png)](https://www.zymoresearch.de/)
[![IIT](https://static.igem.org/mediawiki/2021/e/e0/T--Bielefeld-CeBiTec--iit-logo.jpg)](https://www.iit-biotech.de/)
![Implen](https://static.igem.org/mediawiki/2021/9/93/T--Bielefeld-CeBiTec--implen-logo.jpg)
![JBV](https://static.igem.org/mediawiki/2021/b/b1/T--Bielefeld-CeBiTec--jbv-logo.png)
[![Zierold-Stiftung](https://static.igem.org/mediawiki/2021/a/a9/T--Bielefeld-CeBiTec--zierold-stiftung-logo.jpg)](https://www.zierold-stiftung.de/)
![roth](https://static.igem.org/mediawiki/2021/3/38/T--Bielefeld-CeBiTec--Logo_Roth.jpg)
![sarstedt](https://static.igem.org/mediawiki/2021/3/39/T--Bielefeld-CeBiTec--Logo_Sarstedt.png)
![dephyte](https://static.igem.org/mediawiki/2021/c/c6/T--Bielefeld-CeBiTec--Logo_Dephyte.jpg)
![microsynth](https://static.igem.org/mediawiki/2021/7/7d/T--Bielefeld-CeBiTec--Microsynth_logo.svg)
![jena-bioscience](https://static.igem.org/mediawiki/2021/7/7e/T--Bielefeld-CeBiTec--Logo_Bioscience.png)
![redbull](https://static.igem.org/mediawiki/2021/9/92/T--Bielefeld-CeBiTec--Logo_redbull.svg)
[![biozym](https://static.igem.org/mediawiki/2021/c/c6/T--Bielefeld-CeBiTec--Logo_biozym.png)](https://www.biozym.com/)
[![Chris Pratt on Camel](https://static.igem.org/mediawiki/2021/5/5a/T--Bielefeld-CeBiTec--Chris_on_Camel.png)](https://www.youtube.com/watch?v=dQw4w9WgXcQ)

[![](https://static.igem.org/mediawiki/2021/8/8b/HQ_page_logo.jpg)](https://2021.igem.org)

# Team:Bielefeld-CeBiTec/Engineering

[![Home](https://static.igem.org/mediawiki/2021/4/4b/T--Bielefeld-CeBiTec--21_PLANT_logo.png)](https://2021.igem.org/Team%3ABielefeld-CeBiTec)

[ ]

People

[Team](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Team)

[Attribution](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Attributions)

[Partners](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Sponsoring)

[Medal Criteria](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Medal_Criteria)

[Bielefeld](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Bielefeld)

Human Practices

[Overview](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Human_Practices_Overview)

[Human Practices](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Human_Practices)

[Education](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Education)

[Experts](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Experts)

[Communication](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Communication)

[Collaborations](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Collaborations)

iGEM Phototrophs

[Partnership](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Partnership)

[Contribution](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Contribution)

Lab

[Experiments](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Experiments)

[Parts](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Parts)

[Basic Part](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Basic_Part)

[Improve-a-Part](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Improve)

[Safety](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Safety)

[Notebook](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Notebook)

Results

[Overview](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Results_Overview)

[Hydroculture](https://2021.igem.org/Team%3ABielefeld-CeBiTec/results_hydroculture)

[Bacteria](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Bacteria)

[Tobacco](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Tobacco)

[Reporter](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Reporter)

Project

[Overview](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Overview)

[Description](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Description)

[Model](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Model)

[Proof of Concept](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Proof_Of_Concept)

[Implementation](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Implementation)

[Engineering](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Engineering)

[Chemicals](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Chemicals)

[Outlook](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Outlook)

![](https://static.igem.org/mediawiki/2021/a/a0/T--Bielefeld-CeBiTec--Engineering_cycle_bacteria.png)

## Engineering

Synthetic biology is a branch of biology in which biological systems are engineered to reach a priorly defined goal. Consequently, engineering principles are applied for an effective development. One of the basic principles is the engineering cycle, that is composed of 4 stages: Design, Build, Test, Learn. Once the topic is selected and extensive research was performed, a conceptualization phase must be conducted before the design cycle is entered. In this phase the overall goals are specified, inputs and outputs are defined. For our project we had 5 main goals.

1. Establishing the signaling cascade in bacteria
2. Optimizing the signaling cascade
3. Testing our computationally designed receptors *in vivo*
4. Optimizing the receptors
5. Establishing the signaling cascade in tobacco

The inputs were defined as the chemical weapon degradation products or simulants, that bind to our computationally designed receptors. The output for bacteria was specified as GFP and for the subsequent integration in tobacco as RUBY or ANTHOS.

To achieve the goals, we used the principles of the engineering cycle. In the initial cycle we started by selecting the genetic parts for our signaling cascade and by designing a single plasmid, that had to be cloned into *E. coli* DH5α in the building stage of the first cycle. Subsequently we tested the signaling cascade by cultivation with glucose and ribose, ligands of the galactose/glucose binding protein (GBP) and the ribose binding protein (RBP) respectively. The receptors were expressed endogenously in *E. coli*. The cultures were analyzed on the GFP fluorescence and confirmed by confocal laser scanning microscopy, the output we defined beforehand. A low but visible fluorescence was detected. The fluorescence output for RBP was higher than for the GBP tests. Higher concentration of additives does not lead to a higher output. From this we learned, that the RBP is a more suitable receptor for subsequent experiments and suspected, that the RBP expression might be a bottleneck in the signaling cascade, because GFP expression did not increase with increasing concentrations of the inductor. With the ending of the first cycle, we reached the goal of establishing the signaling cascade in bacteria.

Since we learned that expression of RBP seems to be a limiting factor, we decided to overexpress it in a second cycle to optimize the signaling cascade and accomplish the second goal. In the design stage, we designed a plasmid for overexpression of RBP. The plasmid was built and co-transformed with the signaling cascade into *E. coli* BL21(DE3). In a cultivation experiment with supplemented ribose as the ligand, we measured the fluorescence and determined, that the fluorescence increased significantly after overexpression of RBP. We learned, that RBP expression was in fact a bottleneck of the signaling cascade. The second goal of optimizing the signaling cascade was therefore reached and the second cycle completed. Furthermore, we obtained clearly differentiable GFP signals, we could base the next experiments on.

The signaling cascade from the first cycle was also cloned into *E. coli* BL21(DE3) and the after a cultivation experiment with ribose added, we measured the fluorescence and found, that the fluorescence increased significantly after overexpression of RBP. We learned, that RBP was in fact a bottleneck of the signaling cascade. The second goal of optimizing the signaling cascade was therefore reached and the second cycle completed. Furthermore, we obtained clearly differentiable GFP signals, we could base the next experiments on.

In the third cycle we tested our computationally designed receptors. We designed two plasmids for overexpression of the BTCA-receptor and the DIMP-receptor, that were also used for expression of receptors for *in vitro* testing. The plasmids were built and co-transformed with the signaling cascade into *E. coli* BL21(DE3). With fluorescence measurements we confirmed that the addition of BTCA resulted in a higher fluorescence of the cultures with the BTCA-receptor overexpression than in the cultures with the RBP overexpression. The overexpression of the DIMP-receptor led to no difference increased fluorescence intensity and therefore could not be shown to
be functional to bind DIMP. 3D deconvolution widefield fluorescent microscopy and super resolution 3D-structured illumination microscopy confirmed that we were in fact measuring GFP fluorescence from the engineered bacteria. From this we learned, that the BTCA receptor seems to have a higher affinity to BTCA than to ribose and a higher affinity to BTCA than RBP. This was a proof of concept for our project, marked the end of the third cycle and the accomplishment of the third goal.

Due to time constraints, we were not able to continue with further cycles. In a fourth cycle we would have redesigned the plasmid from the first cycle and would have integrated the computationally designed receptors. These plasmids would have been constructed and cloned into *E. coli*. Fluorescence measurements could have been performed as before.

The fifth cycle would have been the receptor optimizing cycle. We would either have constructed a library to optimize the existing receptors (e.g.: the DIMP-receptor) or to generate novel receptors. Another approach would have been computational receptor design. The cycle would have been the base for the integration into plants. The first step would have been receptor design by either computational design or the design of primers for a library assembly. The next stage would have been constructing a library with a suitable method like Darwin assembly with subsequent cloning and for the computational design the plasmid construction and cloning. Furthermore, we would have tested the receptors like in the prior cycles and would have screened the output of the signaling cascade. From this we would have drawn conclusions about the quality of the receptors and would either enter the same engineering cycle again or would have integrated the receptor into plants. With this we would have reached our final goal and would have a bacterial based system in which we would have been able to easily design, build and test novel receptors.

---

[![Promega](https://static.igem.org/mediawiki/2021/1/14/T--Bielefeld-CeBiTec--promega-logo.png)](https://www.promega.de/)
[![Merck](https://static.igem.org/mediawiki/2021/1/17/T--Bielefeld-CeBiTec--merck-logo.png)](https://www.merckgroup.com/de)
[![Plasmid-Factory](https://static.igem.org/mediawiki/2021/d/dd/T--Bielefeld-CeBiTec--pf-logo.jpg)](https://plasmidfactory.com/)
[![SnapGene](https://static.igem.org/mediawiki/2021/d/d0/T--Bielefeld-CeBiTec--snapgene-logo.png)](https://www.snapgene.com/)
[![Sparkasse-Bielefeld](https://static.igem.org/mediawiki/2021/5/58/T--Bielefeld-CeBiTec--spkb-logo.png)](https://www.sparkasse-bielefeld.de/de/home.html)
[![Starlab](https://static.igem.org/mediawiki/2021/e/ec/T--Bielefeld-CeBiTec--starlab_logo.svg)](https://www.starlabgroup.com/)
[![Zymo](https://static.igem.org/mediawiki/2021/2/27/T--Bielefeld-CeBiTec--zymo-logo.png)](https://www.zymoresearch.de/)
[![IIT](https://static.igem.org/mediawiki/2021/e/e0/T--Bielefeld-CeBiTec--iit-logo.jpg)](https://www.iit-biotech.de/)
![Implen](https://static.igem.org/mediawiki/2021/9/93/T--Bielefeld-CeBiTec--implen-logo.jpg)
![JBV](https://static.igem.org/mediawiki/2021/b/b1/T--Bielefeld-CeBiTec--jbv-logo.png)
[![Zierold-Stiftung](https://static.igem.org/mediawiki/2021/a/a9/T--Bielefeld-CeBiTec--zierold-stiftung-logo.jpg)](https://www.zierold-stiftung.de/)
![roth](https://static.igem.org/mediawiki/2021/3/38/T--Bielefeld-CeBiTec--Logo_Roth.jpg)
![sarstedt](https://static.igem.org/mediawiki/2021/3/39/T--Bielefeld-CeBiTec--Logo_Sarstedt.png)
![dephyte](https://static.igem.org/mediawiki/2021/c/c6/T--Bielefeld-CeBiTec--Logo_Dephyte.jpg)
![microsynth](https://static.igem.org/mediawiki/2021/7/7d/T--Bielefeld-CeBiTec--Microsynth_logo.svg)
![jena-bioscience](https://static.igem.org/mediawiki/2021/7/7e/T--Bielefeld-CeBiTec--Logo_Bioscience.png)
![redbull](https://static.igem.org/mediawiki/2021/9/92/T--Bielefeld-CeBiTec--Logo_redbull.svg)
[![biozym](https://static.igem.org/mediawiki/2021/c/c6/T--Bielefeld-CeBiTec--Logo_biozym.png)](https://www.biozym.com/)
[![Chris Pratt on Camel](https://static.igem.org/mediawiki/2021/5/5a/T--Bielefeld-CeBiTec--Chris_on_Camel.png)](https://www.youtube.com/watch?v=dQw4w9WgXcQ)

[![](https://static.igem.org/mediawiki/2021/8/8b/HQ_page_logo.jpg)](https://2021.igem.org)

# Team:Bielefeld-CeBiTec/Experiments

[![Home](https://static.igem.org/mediawiki/2021/4/4b/T--Bielefeld-CeBiTec--21_PLANT_logo.png)](https://2021.igem.org/Team%3ABielefeld-CeBiTec)

[ ]

People

[Team](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Team)

[Attribution](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Attributions)

[Partners](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Sponsoring)

[Medal Criteria](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Medal_Criteria)

[Bielefeld](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Bielefeld)

Human Practices

[Overview](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Human_Practices_Overview)

[Human Practices](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Human_Practices)

[Education](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Education)

[Experts](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Experts)

[Communication](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Communication)

[Collaborations](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Collaborations)

iGEM Phototrophs

[Partnership](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Partnership)

[Contribution](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Contribution)

Lab

[Experiments](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Experiments)

[Parts](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Parts)

[Basic Part](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Basic_Part)

[Improve-a-Part](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Improve)

[Safety](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Safety)

[Notebook](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Notebook)

Results

[Overview](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Results_Overview)

[Hydroculture](https://2021.igem.org/Team%3ABielefeld-CeBiTec/results_hydroculture)

[Bacteria](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Bacteria)

[Tobacco](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Tobacco)

[Reporter](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Reporter)

Project

[Overview](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Overview)

[Description](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Description)

[Model](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Model)

[Proof of Concept](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Proof_Of_Concept)

[Implementation](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Implementation)

[Engineering](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Engineering)

[Chemicals](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Chemicals)

[Outlook](https://2021.igem.org/Team%3ABielefeld-CeBiTec/Outlook)

[50x TAE Stock Solution](https://static.igem.org/mediawiki/2021/2/20/T--Bielefeld-CeBiTec--21_TAE_Stock_Solution.pdf)

[Agroinfiltration of *N. Benthamiana* leaves](https://static.igem.org/mediawiki/2021/4/4e/T--Bielefeld-CeBiTec--21_Agroinfiltration.pdf)

[Betalain measurement](https://static.igem.org/mediawiki/2021/0/0c/T--Bielefeld-CeBiTec--21_betalain_mesurement.pdf)

[Colony PCR with Thermo Scientific DreamTaq DNA Polymerase](https://www.thermofisher.com/document-connect/document-connect.html?url=https://assets.thermofisher.com/TFS-Assets%2FLSG%2Fmanuals%2FMAN0012036_DreamTaqDNAPolymerase_500U_UG.pdf)

[Fast cell lysis for SDS-Page](https://static.igem.org/mediawiki/2021/7/71/T--Bielefeld-CeBiTec--21_fast_cell_lysis_SDS_page.pdf)

[GeneJET Plasmid-Miniprep-Kit](https://www.thermofisher.com/document-connect/document-connect.html?url=https://assets.thermofisher.com/TFS-Assets%2FLSG%2Fmanuals%2FMAN0013117_GeneJET_Plasmid_Miniprep_UG.pdf)

[Generating electrocompetent cells](https://static.igem.org/mediawiki/2021/4/4e/T--Bielefeld-CeBiTec--21_electro_purification.pdf)

[Generating Generating chemocompetent cells](https://static.igem.org/mediawiki/2021/e/ef/T--Bielefeld-CeBiTec--21_Aqua_Cloning.pdf)

[Gibson Assembly](https://static.igem.org/mediawiki/2021/f/f3/T--Bielefeld-CeBiTec--21_Gibson_Assembly.pdf)

[Lenox Broth (LB)](https://static.igem.org/mediawiki/2021/a/ad/T--Bielefeld-CeBiTec--21_LB.pdf)

[Minimal Medium](https://static.igem.org/mediawiki/2021/2/27/T--Bielefeld-CeBiTec--21_minimal_medium.pdf)

[Monarch PCR & DNA Cleanup Kit (New England Biolabs GmbH)](https://international.neb.com/-/media/nebus/files/protocols/t1030_quick_protocol_card_monarch_pcrdna_cleanup.pdf?rev=a562d8f1f21741b0ac6d59ea9020cef3&hash=E4845F80E8B2BE925A09DAD6562A41EC)

[PCR with Phusion™ High-Fidelity DNA Polymerase (Thermo Scientific™)](https://www.thermofisher.com/order/catalog/product/de/de/F-530XL)

[GeneJET Plasmid-Miniprep-Kit (Thermo Scientific™)](https://www.thermofisher.com/document-connect/document-connect.html?url=https://assets.thermofisher.com/TFS-Assets%2FLSG%2Fmanuals%2FMAN0013117_GeneJET_Plasmid_Miniprep_UG.pdf)

[SDS Page](https://static.igem.org/mediawiki/2021/f/fb/T--Bielefeld-CeBiTec--21_SDS_Page.pdf)

[SDS Page running buffer](https://static.igem.org/mediawiki/2021/f/fb/T--Bielefeld-CeBiTec--21_SDS-page_running_buffer.pdf)

[SOC Medium](https://static.igem.org/mediawiki/2021/9/92/T--Bielefeld-CeBiTec--21_SOC.pdf)

[TAE Buffer](https://static.igem.org/mediawiki/2021/d/d9/T--Bielefeld-CeBiTec--21_TAE_buffer.pdf)

[Touchdown PCR](https://static.igem.org/mediawiki/2021/7/7c/T--Bielefeld-CeBiTec--21_Touchdown_PCR.pdf)

[Transformation via elektroporation](https://static.igem.org/mediawiki/2021/4/4e/T--Bielefeld-CeBiTec--21_electro_purification.pdf)

[Transformation via heat shock](https://static.igem.org/mediawiki/2021/2/25/T--Bielefeld-CeBiTec--21_heat_shock.pdf)

[“Typical“ Restriction Digest](https://static.igem.org/mediawiki/2021/8/81/T--Bielefeld-CeBiTec--21_Restriction_digest.pdf)

[YEP Medium](https://static.igem.org/mediawiki/2021/c/c6/T--Bielefeld-CeBiTec--21_YEP.pdf)

---

[![Promega](https://static.igem.org/mediawiki/2021/1/14/T--Bielefeld-CeBiTec--promega-logo.png)](https://www.promega.de/)
[![Merck](https://static.igem.org/mediawiki/2021/1/17/T--Bielefeld-CeBiTec--merck-logo.png)](https://www.merckgroup.com/de)
[![Plasmid-Factory](https://static.igem.org/mediawiki/2021/d/dd/T--Bielefeld-CeBiTec--pf-logo.jpg)](https://plasmidfactory.com/)
[![SnapGene](https://static.igem.org/mediawiki/2021/d/d0/T--Bielefeld-CeBiTec--snapgene-logo.png)](https://www.snapgene.com/)
[![Sparkasse-Bielefeld](https://static.igem.org/mediawiki/2021/5/58/T--Bielefeld-CeBiTec--spkb-logo.png)](https://www.sparkasse-bielefeld.de/de/home.html)
[![Starlab](https://static.igem.org/mediawiki/2021/e/ec/T--Bielefeld-CeBiTec--starlab_logo.svg)](https://www.starlabgroup.com/)
[![Zymo](https://static.igem.org/mediawiki/2021/2/27/T--Bielefeld-CeBiTec--zymo-logo.png)](https://www.zymoresearch.de/)
[![IIT](https://static.igem.org/mediawiki/2021/e/e0/T--Bielefeld-CeBiTec--iit-logo.jpg)](https://www.iit-biotech.de/)
![Implen](https://static.igem.org/mediawiki/2021/9/93/T--Bielefeld-CeBiTec--implen-logo.jpg)
![JBV](https://static.igem.org/mediawiki/2021/b/b1/T--Bielefeld-CeBiTec--jbv-logo.png)
[![Zierold-Stiftung](https://static.igem.org/mediawiki/2021/a/a9/T--Bielefeld-CeBiTec--zierold-stiftung-logo.jpg)](https://www.zierold-stiftung.de/)
![roth](https://static.igem.org/mediawiki/2021/3/38/T--Bielefeld-CeBiTec--Logo_Roth.jpg)
![sarstedt](https://static.igem.org/mediawiki/2021/3/39/T--Bielefeld-CeBiTec--Logo_Sarstedt.png)
![dephyte](https://static.igem.org/mediawiki/2021/c/c6/T--Bielefeld-CeBiTec--Logo_Dephyte.jpg)
![microsynth](https://static.igem.org/mediawiki/2021/7/7d/T--Bielefeld-CeBiTec--Microsynth_logo.svg)
![jena-bioscience](https://static.igem.org/mediawiki/2021/7/7e/T--Bielefeld-CeBiTec--Logo_Bioscience.png)
![redbull](https://static.igem.org/mediawiki/2021/9/92/T--Bielefeld-CeBiTec--Logo_redbull.svg)
[![biozym](https://static.igem.org/mediawiki/2021/c/c6/T--Bielefeld-CeBiTec--Logo_biozym.png)](https://www.biozym.com/)
[![Chris Pratt on Camel](https://static.igem.org/mediawiki/2021/5/5a/T--Bielefeld-CeBiTec--Chris_on_Camel.png)](https://www.youtube.com/watch?v=dQw4w9WgXcQ)

[![](https://static.igem.org/mediawiki/2021/8/8b/HQ_page_logo.jpg)](https://2021.igem.org)

# Team:Bielefeld-CeBiTec/Results

# Results

You can describe the results of your project and your future plans here.

### What should this page contain?

* Clearly and objectively describe the results of your work.
* Future plans for the project.
* Considerations for replicating the experiments.

### Describe what your results mean

* Interpretation of the results obtained during your project. Don't just show a plot/figure/graph/other, tell us what you think the data means. This is an important part of your project that the judges will look for.
* Show data, but remember **all measurement and characterization data must also be on the Part's Main Page on the [Registry](http://parts.igem.org/Main_Page).** Otherwise these data will not be in consideration for any medals or part awards!
* Consider including an analysis summary section to discuss what your results mean. Judges like to read what you think your data means, beyond all the data you have acquired during your project.

### Project Achievements

You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.

* A list of linked bullet points of the successful results during your project
* A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.

### Inspiration

See how other teams presented their results.

* [2019 Newcastle](https://2019.igem.org/Team%3ANewcastle/Results)
* [2019 Munich](https://2019.igem.org/Team%3AMunich/Results)
* [2019 Tec Chihuahua](https://2019.igem.org/Team%3ATec-Chihuahua/Results)
* [2020 Aalto Helsinki](https://2020.igem.org/Team%3AAalto-Helsinki/Results)
* [2020 GreatBay SCIE](https://2020.igem.org/Team%3AGreatBay_SCIE/Results)
* [2020 Queens Canada](https://2020.igem.org/Team%3AQueens_Canada/Results)