Team:ZJU-China/Notebook/LabNotes/July

|July 1st-7th Final examinations for the semester.  

|July 8th Transformation of GFP. Pick up two colonies and cultivate in A+LB medium.  

|July 9th Set up Ampicillin (100 mg/mL) and store in -20°C (for future use).  

|July 10th Transform BBa_K411003 into BC21 cells. Transformation of GFP, 23L and theophylline using 100μL/mL Ampicillin.  

|July 11th Observe after ~20h cultivation and find colonies. Pick up colonies and analyze with PCR. Also transform 23L, theophylline and GFP into DH10β cells.  

|July 12th Find that cells with GFP and theophylline parts are in good condition. PCR amplification of Phi X 174 and gene E.  

|July 13th Plasmid pBADS purification. 3A assembly of parts pBADS, PhiX174-pE, and pSB1k3.  

|July 14th Cut pBAD and pE --- successful! Pick up colonies with pBAD-pE and cultivate. Transform pBAD-pE into BL21.  

|July 15th PCR amplification of RBS BBa_J61100 and double terminator BBa_B0015. 3A assembly of three parts.  

|July 16th Cut BBa_K411003 and pSB1A3 with E+P. Link BBa_K411003 with pSB1A3.  

|July 17th Plasmid purification of pE-pSB1c3. Transform GFP and 23L, but no results with 23L. PCR analysis of pBADS-RBS.  

|July 18th Plasmid purifications of pE-DTer, pBAD-RBS. PCR amplification of BBa_J611007.  

|July 19th 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.  

|July 20th Small-scale purification of plasmid with BBa_K411003, and then cut with E+P. Transform Lac I into DH10β cells.  

|July 21st 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).  

|July 22nd Transformation of our last year’s parts into 23L cells --- successful! Pick up colonies and cultivate.  

|July 23rd Measure the static performance of BBa_K411003 in BL21 cells; the results are bad. Purify plasmids 4P, 4B and 6D again.  

|July 24th 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.  

|July 25th Measure the static performance of theophylline. Purification of R0010 plasmid (with Amp or Chl resistance).  

|July 26th PCR amplification to analyze the results of transformation. Cut GFP with E+P again.  

|July 27th Pick up colonies of Streptavidin, and do some purification works.  

|July 28th Cut pBADS-RBS + pE-Ter with E+P. Pick up colonies of parts GTP, B0015, and theophylline.  

|July 29th Link pSB1c3 with linker Streptavidin. Measure static performance of theophylline, and find the OD value rises sharply.  

|July 30th PCR cleanup. Transformation of GFP.  

|July 31st PCR and gel electrophoresis to analyze results of GFP transformation.