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

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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:

- It could measure both absorbance and fluorescence
- It had pathlength correction
- Temperature settings went from 4 to 65°C with a precision of ± 0.5°C at 37°C.
- Bandpass width was 530/20
- Excitation was 485 nm
- It used top optics

0. Transformation of competent cells

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Luckily, we did not need to create our own competent cells as we already had high-efficiency dh5α cells at hand. As we had a sufficient amount of colonies on our plates, we picked two colonies from each plate and inoculated them in LB medium + chloramphenicol O/N at 37°C and 220 rpm.