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Team:HokkaidoU Japan/NotebookTest
From 2010.igem.org
since 13 Jul 2010
since 1 Aug 2010
Calendar
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Abstract
Monday, July 12, 2010
- Our first experiment, transformation of BioBrick devices.
Wednesday, July 21, 2010
- Preparation of competent cells
- Single colony isolation of E. coli which had been transformed on Monday, July 12, 2010
Monday, July 26, 2010
- Cultivation of transformed E. coli for the next day's miniprep
Tuesday, July 27, 2010
- Miniprep
Monday, August 2, 2010
- 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
Monday, August 9, 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 E. coli and heat-sensitive E. coli
- These are preliminary experiments before starting the main project
Tuesday, August 10, 2010
- 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 to confirm
Wednesday, August 11, 2010
- Preparation for the next day's miniprep
- Estimation of enzyme activity
- Preparation of the glycerol stocks
Thursday, August 12, 2010
- Miniprep (1-18F, 2-21H and 2-11P)
- Electrophoresis of the DNA which had been amplified via PCR and miniprep
Friday, August 13, 2010
- PCR amplification of the DNA that we couldn't obtain via miniprep previous day(1-18F)
Monday, August 16, 2010
- Measurement of restriction enzyme activity
- Restriction enzyme digestion of the parts(3-1E, 1-2M, 1-18F and 1-23L) and the vector(pSB1C3)
Tuesday, August 17, 2010
- Purification of the DNA solutions via gel extraction
- Preparation of agar medium containing 35 ug/mL chloramphenicol
Wednesday, August 18, 2010
- Digestion and gel extraction of 1-2M (retry)
- 3 piece ligation of 1-18F, 1-23L and pSB1C3
- Transformation
Thursday, August 19, 2010
- 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
Friday, August 20, 2010
- PCR amplification of pSB1A3, pSB1C3 and pSB1T3
Monday, August 23, 2010
- PCR amplification of pSB1A3, pSB1C3 and pSB1T3 (with some changes in protocols)
- PCR amplification of BioBrick parts using new primers
Tuesday, August 24, 2010
- Electrophoresis of PCR products that had been amplified using new primers
- Examination of two ligation kits
- Electrophoresis assay of miscellaneous DNA solutions
- Transformation of 1-3A (RFP reporter with chloramphenicol resistance) to evaluate the medium
Wednesday, August 25, 2010
- Estimation of the amount of 1-3A DNA that could transform and grow cells on the chloramphenicol medium
- Ethanol precipitation to condense the pSB1C3 DNA solution, and its digestion and ligation
Ligationはお互いそれともパーツと？
ベクター同士．ご自分の記録をご確認ください．
- Digestion of PCR products that had been amplified by using new primers
Thursday, August 26, 2010
- Electriphoresis to check whether ligation had been successful
- Digestion of PCR products that had been amplified using new primers (with reconstruction of reaction system)
Friday, August 27, 2010
- Ligation of 1-1A (RFP reporter device) into pSB1C3 and its transformation
- Retry of 3 piece ligation which was done on August 19th and 20th
- Ligation of vectors to each other
Monday, August 30, 2010
- Measurement of restriction enzyme activity using highly purified pUC119
- Digestion of PCR products that had been amplified by using new primers
amplified using
Tuesday, August 31, 2010
- Gel extraction, ligation and transformation of pUC119
Wednesday, September 1, 2010
- PCR amplification of 1-5A (RFP reporter)
Thursday, September 2, 2010
- Digestion, ligation and Transformation of RFP reporter with pSB1C3 or pUC119
Friday, September 3, 2010
- Colony PCR of previous day's transformed E. coli
Colony PCR of ''E. coli'' transformed on previous day
- PCR amplification of 1-3A (RFP reporter)
Saturday, September 4, 2010
- PCR amplification of 1-5A (RFP reporter)
- Estimation of the amount of 1-3A PCR product
