Calendar
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Abstract
- The first experiment for us, transformation with BioBrick Devices.
- Preparation of competent cells
- Single colony isolation of E. coli which had been transformed on Monday, July 12, 2010
- Cultivation of transformed E. coli for the next day's miniprep
- Restriction enzyme digestion of plasmids which had been purified by miniprep
- Electrophoresis assay
Tuesday, August 3, 2010
Wednesday, August 4, 2010
Thursday, August 5, 2010
Friday, August 6, 2010
- Initial amplification of the BioBrick parts (BBa_I14032 "2-11P", BBa_F2621 "2-21H", BBa_K098995 "3-1E" and BBa_E1010 "1-18F") that are used in the construction of concentration-sensitive and heat-sensitive E. coli
- This is a preliminary experiment before starting the main project
- PCR amplification of the other parts (BBa_F1610 "2-24G", BBa_B0034 "1-2M", BBa_B0015 "1-23L" and BBa_K098995 "3-1E") and electrophoresis for confirmation
- Preparation for the next day's miniprep
- Estimation of enzyme activity
- Preparation of the glycerol stocks
- Miniprep (1-18F, 2-21H and 2-11P)
- Electrophoresis of the DNA which had amplified via PCR and miniprep
- PCR amplification of the DNA that we couldn't obtain via miniprep previous day(1-18F)
- Measurement of restriction enzyme activity
- Restriction enzyme digestion of the parts(3-1E, 1-2M, 1-18F and 1-23L) and the vector(pSB1C3)
- Purification of the DNA solutions via gel extraction
- Preparation of agar medium containing 35 ug/mL chloramphenicol
- Digestion and gel extraction of 1-2M (retry)
- 3 piece ligation of 1-18F, 1-23L and pSB1C3
- Transformation
- 3 piece ligation of 3-1E (heat sensor), 1-2M (ribosome binding site) and pSB1C3 (vector)
- Ligation that uses only vector pSB1C3 to estimate its efficiency
PCR amplification of pSB1A3, pSB1C3 and pSB1T3
===
Tuesday, November 30, 2010===