Team:UChicago/Notebook

June

Week 3

Tuesday, June 18, 2013

First iGEM Meeting of the Summer!

• All student members gathered in the lab.

• We toured the lab, prep room, and incubator room, and we discussed the lab rules.

• We also split into teams to discuss lab logistics: contacting funding sources, ordering reagents/materials, compiling protocols, recording our electronic lab notebook, and making plates/media.

Week 4

Wednesday, June 26, 2013

Planning Wet Lab Work

• We split into planning sub-teams to plan out different parts of the project with protocols, timelines, and ordering reagents.
  • Plasmid Design
  • Media and Competent Cells
  • E. coli and B. subtilis Transformation
  • Keratinase Assay and His tagging
  • Running DNA gels, SDS-PAGE gels, and western blots
• We will also be holding weekly meetings with our graduate student advisers and biweekly meetings with our faculty advisers to provide updates.
• Wet lab work begins tomorrow!

Friday, June 28, 2013

Researching the Sequence of kerA

• Planning subteams continued to order reagents and compile protocols.
• The plasmid design team decided on gibson assembly to produce the kerA biobrick with geneblocks ordered from IDT.
• kerA sequence was obtained from “Nucleotide Sequence and Expression of kerA, the Gene Encoding a Keratinolytic Protease of Bacillus licheniformis PWD-1” (Lin et al. 1995).

• Two putative promoters (red), a possible ribosomal binding site (green), and a transcriptional terminator sequence (light blue, single underline) are indicated. The putative starting residues of the preprotein (pre = blue), proprotein (pro = pink), and mature protein (mature = purple) are indicated.
• EcoRI (orange) and PstI (blue) sites within the gene are indicated and must be removed to produce the kerA biobrick.

July

Week 1

Week 2

Week 3

Week 4

August

Week 1

Week 2

Week 3

Week 4

September

Week 1

Week 2

Week 3