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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=== Part 2 ===
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!Date!!Notes
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|-
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=== The Experiments ===
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|July 1st-7th Final examinations for the semester.
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|-
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=== Part 3 ===
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|July 8th Transformation of GFP. Pick up two colonies and cultivate in A+LB medium.
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|-
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== Results ==
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|July 9th Set up Ampicillin (100 mg/mL) and store in -20°C (for future use).
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|-
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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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|-
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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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|-
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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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|-
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|July 13th Plasmid pBADS purification. 3A assembly of parts pBADS, PhiX174-pE, and pSB1k3.
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|-
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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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|-
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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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|-
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|July 16th Cut BBa_K411003 and pSB1A3 with E+P. Link BBa_K411003 with pSB1A3.
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|-
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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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|-
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|July 18th 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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|-
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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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|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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|-
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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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|-
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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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|-
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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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|-
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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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|July 26th 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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|-
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|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.
 +
|-
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|July 30th 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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|}
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Revision as of 15:53, 25 September 2013

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Lab Notes: July

DateNotes
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.

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