Week 1 (4-10 July)

• Lab safety training
• Lab sorting
• Growth Media preparation

Week 2 (11-17 July)

• Primers receive and preparation
• Chemically competent E.coli and Agrobacterium Tumefaciens preparation

Week 3 (18-24 July)

• iGEM 2021 Distribution Kit troubleshooting

Week 4 (25-31 July)

• OmpA PCR amplification 
• Gel electrophoresis of OmpA PCR product and DNA purification

Week 5 (1-7 August)

• PCR amplification of LRR, NB-LRR and Nsm^115-135
• Gel electrophoresis for LRR, NB-LRR and Nsm^115-135

Week 6 (8-14 August)

• Golden Gate Cloning to assemble pLCH86988:OmpA:LRR and pLCH86988:OmpA:NB-LRR
• Transformation of E.coli cells
• Plasmid DNA extraction and purification

Week 7 (22-28 August)

• Diagnostic restriction enzyme digestion and gel electrophoresis

Week 8 (29 August - 4 September)

• Golden Gate Cloning to assemble pLCH86988:Nsm^115-135:YFP
• Transformation of Agrobacterium Tumefaciens cells

Week 9 (5-11 September)

• Agroinfiltration of Agrobacterium Tumefaciens into N. Sylvestris leaves
• Protein extraction from N. Sylvestris leaves

Week 10 (12-18 September)

• Colony PCR
• PCR for OmpA:LRR and OmpA:NB-LRR

Week 11 (19-25 September)

• Cloning with restriction enzymes
• Cloning product clean-up

Week 12 (26 September - 2 October)

• Transformation of E.coli cells to surface display the constructs
• Plasmid DNA extraction and purification
• Restriction enzyme digestion

Week 13 (3-9 October)

• Final Experiment