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Team:Davidson-MissouriW/Notebook
From 2010.igem.org
Notebook
June 1, 2010
- Tested to find lethal concentration of Tet in plated and liquid mediums
- Verified RCBS with our sequences and researched codon optimization methods
- Analyzed probabilities for constructs A and B
June 2, 2010
- Mini prepped, diagnostic RP digest, preparative digest, ligation, pBAD+RBS-RFP
- Researched different ways to optimize a gene
- Began simulation for Constructs A and B
June 3, 2010
- Mini prepped, diagnostic RP digest, preparative digest, ligation, pBAD+RBS-RFP
- Researched restriction sites and created proposal for gene optimization
- Continued work on simulation in Java and Matlab
June 4, 2010
- Mini prepped RBS-TetA-TT
- RX Fragment digestion for RBS-TetA-TT
June 7, 2010
- Ligate and plasmid check for pSB1A2 or pSB1AK3
June 8, 2010
- Double digest on pBAD-RBS-RFP to make an RS fragment insert.
June 9, 2010
- EcoRI and PstI digest on pLac and pBad colonies.
- Arabinose and IPTG concentration experiments started.
June 10, 2010
- Picked 14 colonies from pLac-RBS-RFP and pLac-RBS-RFP-RBS plates.
- Started inducer curve experiments.
June 11, 2010
- Ligated S03736 into pSB1A7 vector.
- EcoRI/PstI digest on pLac-RBS-RFP-RBS colonies.
June 14, 2010
- EcoRI/PstI diagnostic digest performed on pBad-RBS-RFP colonies.
- Inducer curve experiment started on pBad and pLac constructs.
June 15, 2010
- Ligated pLac-RBS-RFP + RBS-TetA.
- IPTG experiment started on pLacI-RBS-RFP
June 16, 2010
- EcoRI/PstI diagnostic digest performed on pLac-RBS-RFP in pSB1A7 vector.
- IPTG experiment started on plates with pLac and pLacI constructs.
June 17, 2010
- Flourimeter data collected from IPTG experiment.
- Ligated pLacI-RBS-RFP + RBS-TetA.
June 18, 2010
- Picked 4 colonies from pLacI-RBS-RFP-RBS-TetA plates.
- IPTG experiment started using newly created IPTG.
June 21, 2010
- Digested TetA with EcoRI/NheI.
- Annealed oligos for optimized and deoptimized TetA segment 1.
June 22, 2010
- Ligated annealed TetA oligos into digested TetA.
- Inducer curve experiment started on pLac and pLacI constructs.
June 23, 2010
- Preparative NheI/BamHI digest performed on TetA.
- Preparative EcoRI/NheI digest performed on TetA.
June 24, 2010
- Annealed optimized and deoptimized segment 1 TetA ligated into vector.
- Annealed optimized and deoptimized segment 2 TetA ligatied into vector.
June 27, 2010
- Inducer curve experiment started on pLac and pLacI constructs.
- Picked colonies from TetA segment 1 & 2 optimized & deoptimized plates.
June 29, 2010
- Screened segment 1 optimized and deoptimized candidate clones using RFLP
- Screened segment 2 optimized and deoptimized candidate clones using RFLP
June 30, 2010
- Tet titration experiment performed on pLac and pLacI +RBS-RFP-RBS-TetA.
- Ligated TetA onto pLac and pLacI +RBS-RFP-RBS-TetA with no terminator.
July 1, 2010
- Preparative BamHI/NheI digest performed on TetA.
- New experiment protocol developed for reduced vector background noise.
July 2, 2010
- Mini prepped, diagnostic RP digest, preparative digest, ligation, pBAD+RBS-RFP
July 6, 2010
- Ligated S04446 and S04447 to J31007.
- Tet titration experiment performed on pLac and pLacI +RBS-RFP-RBS-TetA.
July 7, 2010
- Ligated S04448 to RBS-TetA.
- Ligated S04449 to RBS-TetA.
July 8, 2010
- IPTG experiment started using pre-induced method.
- IPTG experiment started using varying levels of tetracycline.
July 9, 2010
- Picked optimized and deoptimized TetA colonies.
- Flourimeter data collected from IPTG experiment.
July 10, 2010
- Used RFLP to screen for candidate TetA clones.
- Transformed various pLac and pLacI constructs into MG1655 cells.
July 13, 2010
- Used RFLP to screen for candidate TetA clones.
- EcoRI/PstI diagnostic digest performed on K199150.
July 14, 2010
- Ligated segment 2 optimized and deoptimzed TetA behind segment 1.
- IPTG experiment started on I715039-1 and I715039-2.
July 15, 2010
- Cell count data collected from IPTG experiments.
- Flourimeter data collected from IPTG experiments.
July 16, 2010
- Tet titration experiment started using pSB3T5 vector.
- Tet titration experiment started using optimized and deoptimized TetA.
July 19, 2010
- Cell count data collected from Tet titrations.
- Tet titration experiment started using optimized and deoptimized TetA.
June 3, 2010
- Mini prepped, diagnostic RP digest, preparative digest, ligation, pBAD+RBS-RFP
July 20, 2010
- Flourimeter data collected on IPTG experiment.
July 21, 2010
- IPTG experiment started on optimized and deoptmized TetA constructs.
- Passage I715039-1 and I715039-2 experiments started.
July 22, 2010
- IPTG experiment started using varying tetracycline concentrations.
- Cell count data collected on IPTG TetA experiment.
July 23, 2010
- Cell count data collected from IPTG experiment..
- Flourimeter data collected from IPTG experiment.
July 26, 2010
- Presented Missouri Western data to Davidson colleagues.
- Presented Davidson data to Missouri Western colleagues.
July 27, 2010
- Worked on team wiki.
July 28, 2010
- n/a
- n/a
July 30, 2010
- n/a
- n/a
