Team:ZJU-China/Notebook/LabNotes/July

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!Notes
!Notes
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|July 1st-7th||Final examinations for the semester.  
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|Jul 1-7||Final examinations for the semester.  
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|July 8th||Transformation of GFP. Pick up two colonies and cultivate in A+LB medium.  
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|Jul 8||Transformation of GFP. Pick up two colonies and cultivate in A+LB medium.  
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|July 9th||Set up Ampicillin (100 mg/mL) and store in -20°C (for future use).  
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|Jul 9||Set up Ampicillin (100 mg/mL) and store in -20°C (for future use).  
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|July 10th||Transform BBa_K411003 into BC21 cells. Transformation of GFP, 23L and theophylline using 100μL/mL Ampicillin.  
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|Jul 10||Transform BBa_K411003 into BC21 cells. Transformation of GFP, 23L and theophylline using 100μL/mL Ampicillin.  
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|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.  
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|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.  
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|July 12th||Find that cells with GFP and theophylline parts are in good condition. PCR amplification of Phi X 174 and gene E.  
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|Jul 12||Find that cells with GFP and theophylline parts are in good condition. PCR amplification of Phi X 174 and gene E.  
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|July 13th||Plasmid pBADS purification. 3A assembly of parts pBADS, PhiX174-pE, and pSB1k3.  
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|Jul 13||Plasmid pBADS purification. 3A assembly of parts pBADS, PhiX174-pE, and pSB1k3.  
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|July 14th||Cut pBAD and pE --- successful! Pick up colonies with pBAD-pE and cultivate. Transform pBAD-pE into BL21.  
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|Jul 14||Cut pBAD and pE --- successful! Pick up colonies with pBAD-pE and cultivate. Transform pBAD-pE into BL21.  
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|July 15th||PCR amplification of RBS BBa_J61100 and double terminator BBa_B0015. 3A assembly of three parts.  
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|Jul 15||PCR amplification of RBS BBa_J61100 and double terminator BBa_B0015. 3A assembly of three parts.  
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|July 16th||Cut BBa_K411003 and pSB1A3 with E+P. Link BBa_K411003 with pSB1A3.  
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|Jul 16||Cut BBa_K411003 and pSB1A3 with E+P. Link BBa_K411003 with pSB1A3.  
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|July 17th||Plasmid purification of pE-pSB1c3. Transform GFP and 23L, but no results with 23L. PCR analysis of pBADS-RBS.  
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|Jul 17||Plasmid purification of pE-pSB1c3. Transform GFP and 23L, but no results with 23L. PCR analysis of pBADS-RBS.  
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|July 18th||Plasmid purifications of pE-DTer, pBAD-RBS. PCR amplification of BBa_J611007.  
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|Jul 18||Plasmid purifications of pE-DTer, pBAD-RBS. PCR amplification of BBa_J611007.  
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|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.  
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|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.  
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|July 20th||Small-scale purification of plasmid with BBa_K411003, and then cut with E+P. Transform Lac I into DH10β cells.  
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|Jul 20||Small-scale purification of plasmid with BBa_K411003, and then cut with E+P. Transform Lac I into DH10β cells.  
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|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).  
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|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).  
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|July 22nd||Transformation of our last year’s parts into 23L cells --- successful! Pick up colonies and cultivate.  
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|Jul 22||Transformation of our last year’s parts into 23L cells --- successful! Pick up colonies and cultivate.  
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|July 23rd||Measure the static performance of BBa_K411003 in BL21 cells; the results are bad. Purify plasmids 4P, 4B and 6D again.  
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|Jul 23||Measure the static performance of BBa_K411003 in BL21 cells; the results are bad. Purify plasmids 4P, 4B and 6D again.  
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|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.  
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|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.  
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|July 25th||Measure the static performance of theophylline. Purification of R0010 plasmid (with Amp or Chl resistance).  
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|Jul 25||Measure the static performance of theophylline. Purification of R0010 plasmid (with Amp or Chl resistance).  
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|July 26th||PCR amplification to analyze the results of transformation. Cut GFP with E+P again.  
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|Jul 26||PCR amplification to analyze the results of transformation. Cut GFP with E+P again.  
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|July 27th||Pick up colonies of Streptavidin, and do some purification works.  
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|Jul 27||Pick up colonies of Streptavidin, and do some purification works.  
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|July 28th||Cut pBADS-RBS + pE-Ter with E+P. Pick up colonies of parts GTP, B0015, and theophylline.  
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|Jul 28||Cut pBADS-RBS + pE-Ter with E+P. Pick up colonies of parts GTP, B0015, and theophylline.  
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|July 29th||Link pSB1c3 with linker Streptavidin. Measure static performance of theophylline, and find the OD value rises sharply.  
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|Jul 29||Link pSB1c3 with linker Streptavidin. Measure static performance of theophylline, and find the OD value rises sharply.  
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|July 30th||PCR cleanup. Transformation of GFP.  
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|Jul 30||PCR cleanup. Transformation of GFP.  
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|July 31st||PCR and gel electrophoresis to analyze results of GFP transformation.  
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|Jul 31||PCR and gel electrophoresis to analyze results of GFP transformation.  
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Latest revision as of 09:49, 27 September 2013

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Lab Notes: July

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

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