Team:Hong Kong HKU

From 2014.igem.org

(Difference between revisions)
Line 665: Line 665:
                                 <script>
                                 <script>
                                 function myFunction9() {
                                 function myFunction9() {
-
                                     alert("The cloning begins with using two PCR reactions creating the T7 promoter and the signal peptide coding region, and cloned into pET28a using a rolling assembly approach with traditional restriction-ligation cloning. The resulting pET28a containing the desired T7-Signal peptide region was confirmed by sequencing. We then moved on to construct the desired EutSMNLK gene cluster region, which was trouble as a cloning site has to be introduced between the Plac promoter and the EutS gene. Considering the timing and resources, we again used a two-step, traditional, rolling assembly approach, using BBa_K311004 as the template for PCR and pET28a as a platform vector for assembly. Finally, as we confirmed that the desired modified-EutSMNLK region was constructed using colony-PCR test (the binding region for T7Pro sequencing primer was lost in the cloning process due to limitations in restriction enzyme choices), we PCR-ed the whole desired region and ligated into the previous plasmid. The sequence of the plasmid was confirmed by sequencing. For laboratory details about the cloning methodology, please refer to the Methods and Protocols page for reference.");
+
                                     window.open("https://2014.igem.org/Team:Hong_Kong_HKU/plasmidconstruction");
                                 }
                                 }
                                 </script>
                                 </script>

Revision as of 17:31, 16 October 2014

<!DOCTYPE html> HKU IGEM Team

  • Testing
  • Background
  • Products
  • Team
  • Lab results
  • Molecular cloning
  • Protein expression
  • Expression optimization
  • BMC purification
  • Cargo localization
  • BMC-Cargo co-purification
  • Confocal microscopy
  • Transmission electron microscopy
  • Plasmid design
  • Plasmid construction
  • Human practice
  • Card game
  • School tour
  • Biotechnological course
  • Biobrick submission
  • Biobrick characterization
  • Others
  • Safety notes
  • Laboratory notes
  • Methods and Protocols
  • References
  • Acknowledgement

Retrieved from "http://2014.igem.org/Team:Hong_Kong_HKU"