Team:Caltech/Notebook

Notebook

Methods and Protocols

Week 1: 6/15-6/17: Lab orientation

Week 2: 6/19-6/25: Lab training; sample collection, enrichment cultures, DNA extraction from samples

Week 3: 6/26-7/2: Prepare parts by PCR for Gibson assembly of test plasmids of BisdA and BisdB, continue enrichment cultures

Week 4: 7/3-7/10: Sequencing and testing of Gibson assembled vectors

Week 5: 7/11-7/16: Remake Gibson vectors, begin 16S sequencing, run pulsed field gel to extract degradation genes for fosmid kit

Week 6: 7/17-7/23: Continuation of Gibson assembly, 16s sequencing and assembly, and fosmid kit

Week 7: 7/24-7/30: p450 testing, plating of EDC's, more assembly

Week 8: 7/31-8/6: Titering of fosmid kits, HPLC of p450 degradation, more assembly and EDC plating

Week 9: 8/7-8/13 GCMS attempts at p450-degraded BPA, biofilm/crystal violet experiments, assembly of constructs and DDT gene

Week 10: 8/14-8/20: Electrospray of p450-degraded BPA; visits to local water treatment plants; assembly of constructs, lacZ, and DDT gene

Week 11: 8/21-8/27: Assembly of DDT-degradation gene and p450 into BioBrick submission and pET protein expression vectors

Week 12: 8/28-9/3: Biofilm-X-gal experiments, sequencing, HPLC

Week 13: 9/4-9/10: Quikchange, Biobrick, sequencing

Week 14: 9/12-9/18: Redo Quikchange, Biofilm, Protein Expression, HPLC

Week 15: 9/19-9/23: Submit parts to registry: DDT gene, bisdA & bisdB, Wt-F87A