Team:TU Darmstadt/Notebook/sec1/K1602006

• Home
• SLA 3D-Printer
• LabSurfing
• Bricks
• RnD-Nerf

• Sitemap
• Team
• Contact

• PROJECT
  • Biotechnology
    • Monomers
      • Ethylene Glycol
      • Itaconic Acid
      • Xylitol
      • Results
    • In Vitro Degradation
      • In Vitro Bioreactor
      • Xylan Degradation
      • Polyester Degradation
    • Safety
      • Killswitch Design
      • Results
    • Modeling
      • Riboswitch Design
      • Structure Prediction
      • Membrane Modeling
  • Chemistry
  • Engineering
    • Software
    • Hardware
    • Hardening and Printing
  • Policy & Practices
    • LabSurfing
    • Application
    • Techno-moral scenarios
    • Expert talks & Conference
• PARTS
  • Part Collection
• NOTEBOOK
  • Labjournal
  • Methods & Materials
• COLLABORATIONS
  • Testing
  • "Escape the lab"-Tour
  • Open Science
• ATTRIBUTIONS
• TEAM
  • Allocations
  • Project Management in Science
  • Acknowledgements
• GALLERY
  • SLA 3D Printer
  • Lab Impressions
  • Workshops
  • Experts
  • LabSurfing
• SITEMAP

K1602006 - Xylose to Xylitol converting Construct (T7-B0034-GRE3)

Figure 1 cPCR of psB1C3-RBS-GRE3 (X). The size of the amplified product was around 1.4 kbp. DNA marker: 2-Log DNA Ladder (NEB).

To include the RBS-GRE3 brick into an IPTG inducable vector plasmid DNA containing RBS-GRE3 was isolated and cut with XbaI and PstI. The psB1A2 vector containing the T7 promoter was cut with SpeI and PstI and afterwards dephosphorylated. Both restriction products were then ligated with T4-ligase and transformed into Top10 E.coli by heat shock transformation. Afterwards the cells were spread out on an AMP agar plate. Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones which were verified by Sanger sequencing. Afterwards the psB1A2 vector containing the brick (RBS-cadA) was cut with EcoRI and PstI as well as a psB1C3. Both restriction products were then ligated with T4-ligase.