Team:ZJU-China/Notebook/LabNotes/July

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== '''Overall project''' ==
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== Lab Notes: July ==
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Tell us more about your project.  Give us background.  Use this is the abstract of your project.  Be descriptive but concise (1-2 paragraphs)
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== Project Details==
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{|
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|-
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!width="80px"|Date
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!Notes
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|-
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|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.
 +
|-
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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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|Jul 13||Plasmid pBADS purification. 3A assembly of parts pBADS, PhiX174-pE, and pSB1k3.
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|-
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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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|Jul 15||PCR amplification of RBS BBa_J61100 and double terminator BBa_B0015. 3A assembly of three parts.
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|-
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|Jul 16||Cut BBa_K411003 and pSB1A3 with E+P. Link BBa_K411003 with pSB1A3.
 +
|-
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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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|Jul 18||Plasmid purifications of pE-DTer, pBAD-RBS. PCR amplification of BBa_J611007.
 +
|-
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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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|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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|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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=== Part 2 ===
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<html>
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<div style="float:left;">Previous:
 +
  <a href="./June">Lab Notes: June</a>
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</div>
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<div style="float:right;">Next:
 +
  <a href="./August">Lab Notes: August</a>
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</div>
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=== The Experiments ===
 
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=== Part 3 ===
 
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== Results ==
 
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</div>
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   </div>
   </div>

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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