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Team:Warsaw/Journal
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Chemical-based transformation. BioBricks: J06504, J23100, B0034, I712074, E0040, B0015, B0017, E2020, E2030. Failure: E2020. Succeed: J06504, J23100, B0034, I712074, E0040, B0015, B0017, E2030. | Chemical-based transformation. BioBricks: J06504, J23100, B0034, I712074, E0040, B0015, B0017, E2020, E2030. Failure: E2020. Succeed: J06504, J23100, B0034, I712074, E0040, B0015, B0017, E2030. | ||
Isolate plasmid DNA by alkaline lysis. Quality cheked with spectrophotomeric analysis with NanoDrop. | Isolate plasmid DNA by alkaline lysis. Quality cheked with spectrophotomeric analysis with NanoDrop. | ||
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====4th week: 22.07.13 – 26.07.13==== | ====4th week: 22.07.13 – 26.07.13==== | ||
Transformation with isolated pladmids and BioBricks: I74609 and K864100. Isolate plasmid DNA (from I73609 and K864100) with mini-prep kit. Digest and ligation of construct: J23100 and B0034 on plasmid pSB1C3. | Transformation with isolated pladmids and BioBricks: I74609 and K864100. Isolate plasmid DNA (from I73609 and K864100) with mini-prep kit. Digest and ligation of construct: J23100 and B0034 on plasmid pSB1C3. | ||
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====5th week: 29.07.13 – 1.08.13==== | ====5th week: 29.07.13 – 1.08.13==== | ||
Preparing construct: J23100 + B0034 + K864100 on plasmid pSB1A3. Measure glowing fluorescent proteins: sfGTP, sfYFP, sfCFP, sfBFP and SYFP2. Superfolded (sf) forms are made by Warsaw Team. SYFP2 is from parts registry. | Preparing construct: J23100 + B0034 + K864100 on plasmid pSB1A3. Measure glowing fluorescent proteins: sfGTP, sfYFP, sfCFP, sfBFP and SYFP2. Superfolded (sf) forms are made by Warsaw Team. SYFP2 is from parts registry. | ||
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====6th week: 5.08.13 – 9.08.13===== | ====6th week: 5.08.13 – 9.08.13===== | ||
Measure glowing fluorescent proteins. Banking strains with our fluorescent proteins. | Measure glowing fluorescent proteins. Banking strains with our fluorescent proteins. | ||
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====7th week: 12.08.13 – 14.08.13==== | ====7th week: 12.08.13 – 14.08.13==== | ||
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Transformation BioBrick: E2050. | Transformation BioBrick: E2050. | ||
PCR: sfBFP N-terminal, sfYFP N-terminal and mCherry (J06504) C-terminal (succeed). Isolate plasmid DNA with: sfGFP (I73609), mOrange (E2050), sfCFP, sfYFP and sfBFP. | PCR: sfBFP N-terminal, sfYFP N-terminal and mCherry (J06504) C-terminal (succeed). Isolate plasmid DNA with: sfGFP (I73609), mOrange (E2050), sfCFP, sfYFP and sfBFP. | ||
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====8th week: 19.08.13 – 23.08.13==== | ====8th week: 19.08.13 – 23.08.13==== | ||
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Cloning sfCFP N-terminal, mCherry N-terminal, sfYFP N-terminal, sfBFP N-terminal and mCherry C-terminal to pSB1C3 plasmid and inserting it to bacteria. | Cloning sfCFP N-terminal, mCherry N-terminal, sfYFP N-terminal, sfBFP N-terminal and mCherry C-terminal to pSB1C3 plasmid and inserting it to bacteria. | ||
Preparing construct: J23100+B0034+E0040 – failure. | Preparing construct: J23100+B0034+E0040 – failure. | ||
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====9th week: 26.08.13 – 30.08.13==== | ====9th week: 26.08.13 – 30.08.13==== | ||
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<gallery widths=600px heights=800px perrow=1 caption=" | <gallery widths=600px heights=800px perrow=1 caption=" | ||
Verification cloning."> | Verification cloning."> | ||
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File:Klonowanie.jpg|Bla bla | File:Klonowanie.jpg|Bla bla | ||
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</gallery> | </gallery> | ||
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And great success! | And great success! | ||
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PCR: sfGFP N-terminal and GFP C-terminal (various attenuation template DNA) and sfC-terminal (to versions: m6 and m12 to all of superfolded fluorescent proteins). Cloning and inserting it to bacteria. | PCR: sfGFP N-terminal and GFP C-terminal (various attenuation template DNA) and sfC-terminal (to versions: m6 and m12 to all of superfolded fluorescent proteins). Cloning and inserting it to bacteria. | ||
Once again preparing construct: J23100+B0034+E0040. | Once again preparing construct: J23100+B0034+E0040. | ||
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====10th week: 2.09.13 – 7.09.13==== | ====10th week: 2.09.13 – 7.09.13==== | ||
Revision as of 17:32, 7 September 2013
Journal
Genetic lab diary
1st week: 2.07.13 – 5.07.13
Preparation of competent cells for chemical-based transfection. Failure.
2nd week: 8.07.13 – 12.07.13
Succeed preparation of competent cells for chemical-based transfection. Verification with iGEM’s transformation kit. Preparation of competent cells for electroporation. Failure.
3rd week: 15.07.13 – 19.07.13
Chemical-based transformation. BioBricks: J06504, J23100, B0034, I712074, E0040, B0015, B0017, E2020, E2030. Failure: E2020. Succeed: J06504, J23100, B0034, I712074, E0040, B0015, B0017, E2030. Isolate plasmid DNA by alkaline lysis. Quality cheked with spectrophotomeric analysis with NanoDrop.
4th week: 22.07.13 – 26.07.13
Transformation with isolated pladmids and BioBricks: I74609 and K864100. Isolate plasmid DNA (from I73609 and K864100) with mini-prep kit. Digest and ligation of construct: J23100 and B0034 on plasmid pSB1C3.
5th week: 29.07.13 – 1.08.13
Preparing construct: J23100 + B0034 + K864100 on plasmid pSB1A3. Measure glowing fluorescent proteins: sfGTP, sfYFP, sfCFP, sfBFP and SYFP2. Superfolded (sf) forms are made by Warsaw Team. SYFP2 is from parts registry.
6th week: 5.08.13 – 9.08.13=
Measure glowing fluorescent proteins. Banking strains with our fluorescent proteins.
7th week: 12.08.13 – 14.08.13
Transformation BioBrick: E2050. PCR: sfBFP N-terminal, sfYFP N-terminal and mCherry (J06504) C-terminal (succeed). Isolate plasmid DNA with: sfGFP (I73609), mOrange (E2050), sfCFP, sfYFP and sfBFP.
8th week: 19.08.13 – 23.08.13
PCR: sfGFP N-terminal (failure), sfCFP N-terminal (succeed) and mCherry N-terminal (succeed). Gel-out: sfYFP N-terminal, sfBFP N-terminal and mCherry C-terminal. Cloning sfCFP N-terminal, mCherry N-terminal, sfYFP N-terminal, sfBFP N-terminal and mCherry C-terminal to pSB1C3 plasmid and inserting it to bacteria. Preparing construct: J23100+B0034+E0040 – failure.
9th week: 26.08.13 – 30.08.13
 Bla bla |
And great success!
PCR: sfGFP N-terminal and GFP C-terminal (various attenuation template DNA) and sfC-terminal (to versions: m6 and m12 to all of superfolded fluorescent proteins). Cloning and inserting it to bacteria. Once again preparing construct: J23100+B0034+E0040.
10th week: 2.09.13 – 7.09.13
Welcome school! Verification cloning. Unfortunately epic fail. PCR: sfGFP N-terminal, GFP N-terminal and GFP C-terminal. At least success!
