Team:ZJU-China/Notebook/LabNotes/July

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Latest revision as of 09:49, 27 September 2013

Lab Notes

May June July August September

Protocol

Brainstorm

The "Elf"

Background Design Results Perspetives

Lab Notes: July

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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