Team:DTU-Denmark/InterLab

• Project
  • Description
  • Design
  • InterLab
  • Results
  • Choice of Organism
  • amilCP Expression
  • melA Expression
  • nat1 Expression
  • Ganoderma Protoplastation
  • Demonstrate
• Modeling
  • Model
  • Growth Modeling
  • Design of Experiments
  • Modeling the Design
  • Structural Integrity
  • Hardware
• Parts
• Journal
  • Notebook
  • Experiments
• 
• People
  • Team Members
  • Attributions
• Human Practices
  • Human Practices
  • Education and Engagement
• Collaboration
  • Collaborations
  • BioBrick Tutorial
• Safety

Hyphae Hackers

Project

Description Design InterLab Results Choice of Organism amilCP Expression melA Expression nat1 Expression Ganoderma Protoplastation Demonstrate

Modeling

Model Growth Modeling Design of Experiments Modeling the Design Structural Integrity Hardware

Parts Overview Journal

Notebook Experiments

People

Team Members Attributions

Safety Human Practices

Human Practices Education & Engagement

Collaborations

Collaborations BioBrick Tutorial

InterLab

To quote the official Interlab Measurement Study site, “reliable and repeatable measurement is a key component to all engineering disciplines”.

As a first step in our wetlab efforts we decided to embark on the fifth Interlab Measurement Study. In previous studies, it was shown that the variability between different lab measurements could be reduced by GFP fluorescence expression against a known concentration of a fluorescent molecule [minder det for meget om det der står på siden?]. However, the number of cells in the sample also holds a large impact on the variability.

Therefore, this year’s interlab study is about reducing the variability in fluorescence measurements between labs by normalizing to CFU or absolute cell count. To this, we firstly needed a plate reader that is capable of measuring both absorbance and fluorescence. We used the Biotek Synergy MX plate reader. Specifications of this model important to the protocol in use was:

• 21/6/2018
• 22/6/2018
• 26/6/2018
• 2/7/2018
• 3/7/2018
• 4/7/2018
• 5/7/2018

Thursday 21/6/2018

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Competent cells were prepared for the interlab studies using the given the iGem 2018 interlab platereader protocol. We did not need to create our own competent cells as we already had high-efficiency dh5α cells ready.

Friday 22/6/2018

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Today we transformed our prepared competent cells by using the iGem 2018 interlab study transformation.

Tuesday 26/6/2018

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Luquid LB media was prepared using the "Luria-Bertani (LB) Medium Preparation" protocol for the interlab studies in the morning, as we did not have enough for the next step. Interlab is put on hold until we have a platereader.

Monday 2/7/2018

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We picked 2 colonies from each of our transformation plates and inoculated them in the apropriate LB and Chloramphenicol medium. The cells were left to grow overnight at 37 degrees Celsius.

Tuesday 3/7/2018

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Today we finally had the platereader available and continued on the protocol from the 21st of June. We encountered som problems with our fluorescein samples and decided to redo them. We also decided to redo the overnight cultures as we would not have the time to use them today

Wednesday 4/7/2018

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As we finally had all our controls finished, we cpntinued with the "Cell measurement" protocol. Our initial absorbance for our samples were way too high. We then finalized this part of the study and continued with the "Colony Forming Units" protocol.

Thursday5/7/2018

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Data was extracted from our platereader and this, together with the CFU data, was sent off to finalize the interlab study. Another thing on our long list that can be ticked off.

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