| Date
| Notes
|
| Jul 1-7 | Final examinations for the semester.
|
| Jul 8 | Transformation of GFP. Pick up two colonies and cultivate in A+LB medium.
|
| Jul 9 | Set up Ampicillin (100 mg/mL) and store in -20°C (for future use).
|
| Jul 10 | Transform BBa_K411003 into BC21 cells. Transformation of GFP, 23L and theophylline using 100μL/mL Ampicillin.
|
| Jul 11 | Observe after ~20h cultivation and find colonies. Pick up colonies and analyze with PCR. Also transform 23L, theophylline and GFP into DH10β cells.
|
| Jul 12 | Find that cells with GFP and theophylline parts are in good condition. PCR amplification of Phi X 174 and gene E.
|
| Jul 13 | Plasmid pBADS purification. 3A assembly of parts pBADS, PhiX174-pE, and pSB1k3.
|
| Jul 14 | Cut pBAD and pE --- successful! Pick up colonies with pBAD-pE and cultivate. Transform pBAD-pE into BL21.
|
| Jul 15 | PCR amplification of RBS BBa_J61100 and double terminator BBa_B0015. 3A assembly of three parts.
|
| Jul 16 | Cut BBa_K411003 and pSB1A3 with E+P. Link BBa_K411003 with pSB1A3.
|
| Jul 17 | Plasmid purification of pE-pSB1c3. Transform GFP and 23L, but no results with 23L. PCR analysis of pBADS-RBS.
|
| Jul 18 | Plasmid purifications of pE-DTer, pBAD-RBS. PCR amplification of BBa_J611007.
|
| Jul 19 | Small-scale plasmid purification of BBa_K411003. Gel electrophoresis analysis of PCR products. Pick up colonies with GFP and cultivate with rocking bed in 37°C. Purify plasmids with GFP after cultivation.
|
| Jul 20 | Small-scale purification of plasmid with BBa_K411003, and then cut with E+P. Transform Lac I into DH10β cells.
|
| Jul 21 | The Lac I transformation on July 20th was a failure (no colonies); doubt if GE apparatus have problems. Small-scale purification of pBAD_S + RBS (BBa_J611007). Make new competent cells (BL21, DH10β).
|
| Jul 22 | Transformation of our last year’s parts into 23L cells --- successful! Pick up colonies and cultivate.
|
| Jul 23 | Measure the static performance of BBa_K411003 in BL21 cells; the results are bad. Purify plasmids 4P, 4B and 6D again.
|
| Jul 24 | Transformations of R0010 (3H) and R0010 (Amp resistance). Small-scale purification of plasmids in 23L. Cut and link GFP and double terminator. Make Chl solution for future use.
|
| Jul 25 | Measure the static performance of theophylline. Purification of R0010 plasmid (with Amp or Chl resistance).
|
| Jul 26 | PCR amplification to analyze the results of transformation. Cut GFP with E+P again.
|
| Jul 27 | Pick up colonies of Streptavidin, and do some purification works.
|
| Jul 28 | Cut pBADS-RBS + pE-Ter with E+P. Pick up colonies of parts GTP, B0015, and theophylline.
|
| Jul 29 | Link pSB1c3 with linker Streptavidin. Measure static performance of theophylline, and find the OD value rises sharply.
|
| Jul 30 | PCR cleanup. Transformation of GFP.
|
| Jul 31 | PCR and gel electrophoresis to analyze results of GFP transformation.
|
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