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Team:TorontoMaRSDiscovery/Notebook/October
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!align="center"|[[Team:TorontoMaRSDiscovery/Team|The Team]] | !align="center"|[[Team:TorontoMaRSDiscovery/Team|The Team]] | ||
!align="center"|[[Team:TorontoMaRSDiscovery/Project|The Project]] | !align="center"|[[Team:TorontoMaRSDiscovery/Project|The Project]] | ||
| - | !align="center"|[[Team:TorontoMaRSDiscovery/Parts| | + | !align="center"|[[Team:TorontoMaRSDiscovery/Parts|BioBricks]] |
!align="center"|[[Team:TorontoMaRSDiscovery/Modeling|Modeling]] | !align="center"|[[Team:TorontoMaRSDiscovery/Modeling|Modeling]] | ||
!align="center"|[[Team:TorontoMaRSDiscovery/Bioinformatics|Bioinformatics]] | !align="center"|[[Team:TorontoMaRSDiscovery/Bioinformatics|Bioinformatics]] | ||
| + | !align="center"|[[Team:TorontoMaRSDiscovery/Safety|Safety]] | ||
!align="center"|[[Team:TorontoMaRSDiscovery/Notebook|Notebook]] | !align="center"|[[Team:TorontoMaRSDiscovery/Notebook|Notebook]] | ||
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=October 1, 2009= | =October 1, 2009= | ||
Revision as of 16:45, 20 October 2009
| Home | The Team | The Project | BioBricks | Modeling | Bioinformatics | Safety | Notebook |
|---|
Contents |
October 1, 2009
- Miniprepped Tet, 5, EncY, C
- Ran minipreps and K plasmid on a gel
- Besides Tet, all samples looked fine
- Poured K plates
October 2, 2009
- Digested K, EncY and ran on gel
October 3, 2009
- Digested EncY and old Tet plasmid -> ran on gel
- EncY looked fine, but Tet did not show up on gel
- Started overnight culture of Tet
October 4, 2009
- Miniprepped, digested Tet and ran on gel
- Tet was confirmed
- gel extracted EncY, Tet and got low yields
- Started overnight ligations
- 1: Digested Tet + EncY + 5
- 2: Gel extraced Tet + Digested EncY + 5
- negative controls were Tet plasmid alone
October 8, 2009
October 9, 2009
- Miniprepped EncY overnight and digested with E,P
- Ran digest on a gel
- Enc part confirmed
- Transfected Enc+5 ligations from Sunday/Monday into competent cells from Invitrogen
- Used NEB Hight Efficiency Transformation Protocol C2987
- did one 10-fold dilution
- transformed 5ul of DNA
- spread 100ul of cells on plate
- shaker/incubator speed 232rpm
- Used NEB Hight Efficiency Transformation Protocol C2987
October 11, 2009
- Checked plates, no colonies yet
- Picked colonies and started overnights from an old 12E plate that wasn't checked: 12E i,ii,iii
October 13, 2009
- 12E i and iii showed growth -> miniprepped
- Digested and ran on 1% agarose gel
- Stained gel for 1 hour in EtBr+buffer
- A faint ~700 band was observed! This could possibly be the 1+2+Enc insert that we want.
- To confirm the insert, we decided to sequence it
October 15, 2009
- Primers for sequencing 12E insert was designed and ordered
October 16, 2009
- EncY prepped and ready to ship down to MIT to submit part to registry
