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Team:ZJU-China/Notebook/LabNotes/August
From 2013.igem.org
Lab Notes: August
| Date | Notes |
|---|---|
| Aug 2 | Link GFP with S+P. PCR amplification of pE-DTer. Small-scale plasmid purification of LuxR and LuxT. |
| Aug 3 | Clean the refrigerators. Pick up three colonies of CheZ. Find out that double terminator B0015, promoter R0010, and BBa_K411003 are usable. |
| Aug 4 | Preparation for iGEM Conference. |
| Aug 5-8 | iGEM Conference @ NCTU, Hsinchu, Taiwan. |
| Aug 9 | Set up culture plates with Chl. Cut and link FA, FB, BLF1, BLF2. |
| Aug 10 | Observe only one colony with FA; analyze the cause of the failure. Set up culture plates with Chl and restore in 4°C. Cut BBa_K411003 and pSB1A3 with Xba I and Spe I and then do the gel electrophoresis. Small-scale plasmid purification with FA. |
| Aug 11 | Sterilization of culture plates and tips on super-clean benches. Blank culture to investigate the source of pollution. Make up ampicillin, kanamycin, spectinomycin, and chloramphenicol again. |
| Aug 12 | Cut pBAD-S and pE-DTer with S+P and then link. |
| Aug 13 | Transform three parts: BBa_I13500, BBa_R0062, and BBa_K081014. |
| Aug 14 | Measure concentrations of plasmids of yesterday’s transformation. |
| Aug 15 | Cut and link BBa_I13500, BBa_R0062, and BBa_K081014. Set up culture plate with Chl. Make up LHB medium. |
| Aug 16 | Cut FA/FB/BLF1/BLF2 with E+P. Small-scale plasmid purification and gel electrophoresis. Sterilize the tips. |
| Aug 17 | Find that the transformation did on Aug 16th was successful, and then pick up two tubes. Cut and link CheZ and luxR. |
| Aug 18 | Cut CheZ, BLF1, BLF2, and pSB1C3 with X+S. Link CheZ and BLF1, BLF2. |
| Aug 19 | Extraction of the genome of Escherichia coli. Pick up monoxenie. Small-scale plasmid purification. |
| Aug 20 | Prepare another genome of E. coli. |
| Aug 21 | Receive a Bacillus subtilis 168 plate, and culture in 37°C. |
| Aug 22 | Make up CheZ competent cell with TSS and CaCl2 methods. PCR amplification of pSB1C3, atrazine riboswitch, che Z, RBS+GFP, and pSB1C3 --- failed. Small-scale plasmid purification of lux I. |
| Aug 23 | Redo yesterday’s PCR work. Pick up three tubes of monoxenie with LB+Amp. Set up five culture plates with LB medium, three culture plates with Chl. |
| Aug 24 | Link cheZ and RFP. Extract E. coli genomic unit. |
| Aug 25 | Link PET and lux I. Sterilize the tips. |
| Aug 26 | PCR amplification of lgt --- successful. Link cheZ and lux R. |
| Aug 27 | PCR amplification of DsbA, lgt2, and lgt-RAW. Cut cheZ with X+P, luxR with S+P. |
| Aug 28 | Receive new strain from Utah, and culture in 37°C. Pick up colonies with holin, antiholin, theo-ribo, Atr-ribo, lgt1, and lgt2. Link luxR and cheZ. |
| Aug 29 | Receive new strain from CAS in Shanghai, with quorum sensing plasmids in it. Set up seven culture plates with kanamycin. Pick up colonies with cheZ (Wuhan), cheZ (Utah), and BacillusI 168, and measure OD600. |
| Aug 30 | Find that the colonies with QS plasmids are in good condition. Pick up two tubes of monoxenie with QS plasmids and do PCR works. |
| Aug 31 | PCR amplifications of parts R0010, BBa_K411003, srvep-backbone, and pSB1C3. |
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