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Team:ZJU-China/Notebook/LabNotes/September
From 2013.igem.org
Lab Notes: September
| Date | Notes |
|---|---|
| Sept 1 | PCR amplifications of lgt, DsbA-BLF1, DsbA-BLF2, addA stand, pSB1C3 linear backbone, GFP double terminator, and atrazine stand. |
| Sept 2 | Cut BBa_K411003 with E+P. Make up new competent cells with LB and glycerin. Transformation with parts in iGEM kits (BBa_K091111, BBa_K091112 in 2012 distribution, and BBa_K091112 in 2011 distribution). |
| Sept 3 | PCR amplification of theo-stand, atw-stand, DsbA-FA-lgt, DsbA-FB-lgt, and GFP with terminator. |
| Sept 4 | Find that the transformation of lacIq and placIQ1 are both successful. Receive plasmids with TrzN degradation gene from NJU. |
| Sept 5 | PCR amplification of RFP, antiholin, Trz N, holing, BLF1, and BLF2. Small-scale plasmid purification of cheZ and pSB1A3. |
| Sept 6 | Small-scale plasmid purification of FA, ribo-cheZ, and atr riboswitch. Cut PE and kanamycin with X+P. |
| Sept 7 | PCA of FB, BLF1, BLF2, and cheZ-RFP-antiholin. CPEC of DsbA-FA-lgt. Direct CPEC of DsbA-FA-lgt, DsbA-FB-lgt, DsbA-BLF1-lgt, DsbA-BLF2-lgt, and cheZ-RFP-antiholin. |
| Sept 8 | PCR amplification of lgt and DsbA-BLF1. CPEC of ptactamaze-lgt complex. |
| Sept 9 | PCR of holing, BLF2, theo, lact, and FB. |
| Sept 10 | Pick up and test four colonies of the UC Davis strain, and inoculate to LB medium. PCR amplification of BLF1-pSB1C3 and FB. |
| Sept 11 | New method: cover E. coli with calcium carbonate and calcium acid phosphate shell (safety issues). Find that E. coli in such shell can express GFP. PCR verification of anti-Dter, BLF1, BLF2. |
| Sept 12 | Redo some steps of yesterday’s PCR. |
| Sept 13 | Cut and link pluxR and cheZ; PCR clean-up. Small-scale plasmid purification of holin and K (mutant). Cut RFP with E+X, pluxR-cheZ with E+S. |
| Sept 14 | PCR amplification of BLF1, PCA-theo, and PCA-addA. Cut pSB1CB and BLF1 with E+B. |
| Sept 15 | Link pluxR-cheZ-RFP and BLH-pSB1C3. Transform addA, FA, FB, BLF1, BLF2, β-tactamase, and BLF1-1C3. Help UC Davis do transform works of three promoters in 2012 kit (BBa_J23108, BBa_J23109, and BBa_J23111). |
| Sept 16 | Pick up monoxenie of B. subtilis and inoculate. |
| Sept 17 | Find that the transformation of 2012 kit was failed. 3A assembly of luxR-cheZ, RFP, and pSB1A3. |
| Sept 18 | Cut II with N+X, III with N+P. PCR amplification of cheZ, I, lgt, and lgt-RAW. |
| Sept 19 | Mid-autumn festival. One-day holiday! |
| Sept 20 | PCR amplification of lgt-RAW, BBa_K411003, lgt-S, and shrep-back. |
| Sept 21 | Pick up five tubes of colonies and do PCR. Run gel, take photo (on computer). Transform K into BL21 (kanamycin resistance). Transform three parts again for UC Davis. |
| Sept 22 | Pick up two tubes of Theo Ribo, Atra Ribo, and A+ each and put in shaker in 37°C. PCR amplification of FA, FB, I, and lgt-RAW. |
| Sept 23 | Redo some steps of PCR yesterday. |
| Sept 24 | Find that the transformation of FB2 was failed, and FB1 successful. Cut to verify FB1 and lgB1. |
| Sept 25 | Measure the concentration curve of E. coli. Make up new Amp and Chl solution. Pick up three tubes of colonies with holin. |
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Lab Notes: August
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Protocol
