Team:ZJU-China/Notebook/LabNotes/August

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Lab Notes

May June July August September

Protocol

Brainstorm

The "Elf"

Background Design Results Perspetives

Lab Notes: August

Date	Notes
Aug 2nd	Link GFP with S+P. PCR amplification of pE-DTer. Small-scale plasmid purification of LuxR and LuxT.
Aug 3rd	Clean the refrigerators. Pick up three colonies of CheZ. Find out that double terminator B0015, promoter R0010, and BBa_K411003 are usable.
Aug 4th	Preparation for iGEM Conference.
Aug 5th-8th	iGEM Conference @ NCTU, Hsinchu, Taiwan.
Aug 9th	Set up culture plates with Chl. Cut and link FA, FB, BLF1, BLF2.
Aug 10th	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 11th	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 12th	Cut pBAD-S and pE-DTer with S+P and then link.
Aug 13th	Transform three parts: BBa_I13500, BBa_R0062, and BBa_K081014.
Aug 14th	Measure concentrations of plasmids of yesterday’s transformation.
Aug 15th	Cut and link BBa_I13500, BBa_R0062, and BBa_K081014. Set up culture plate with Chl. Make up LHB medium.
Aug 16th	Cut FA/FB/BLF1/BLF2 with E+P. Small-scale plasmid purification and gel electrophoresis. Sterilize the tips.
Aug 17th	Find that the transformation did on Aug 16th was successful, and then pick up two tubes. Cut and link CheZ and luxR.
Aug 18th	Cut CheZ, BLF1, BLF2, and pSB1C3 with X+S. Link CheZ and BLF1, BLF2.
Aug 19th	Extraction of the genome of Escherichia coli. Pick up monoxenie. Small-scale plasmid purification.
Aug 20th	Prepare another genome of E. coli.
Aug 21st	Receive a Bacillus subtilis 168 plate, and culture in 37°C.
Aug 22nd	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 23rd	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 24th	Link cheZ and RFP. Extract E. coli genomic unit.
Aug 25th	Link PET and lux I. Sterilize the tips.
Aug 26th	PCR amplification of lgt --- successful. Link cheZ and lux R.
Aug 27th	PCR amplification of DsbA, lgt2, and lgt-RAW. Cut cheZ with X+P, luxR with S+P.
Aug 28th	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 29th	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 30th	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 31st	PCR amplifications of parts R0010, BBa_K411003, srvep-backbone, and pSB1C3.