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Virginia/8 July 2008
From 2008.igem.org
(Difference between revisions)
(New page: ==Goals== *Prepare overnight broth of successful single colony of Promoter + RBS transformation *Plate Screening plasmid (psb1a10) when it arrives from iGem *Try again with ligation of RFP...) |
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==Goals== | ==Goals== | ||
| - | *Prepare overnight broth of | + | *Prepare overnight broth of single successful colony of Promoter + RBS transformation |
*Plate Screening plasmid (psb1a10) when it arrives from iGem | *Plate Screening plasmid (psb1a10) when it arrives from iGem | ||
*Try again with ligation of RFP and GFP enzyme cut DNA | *Try again with ligation of RFP and GFP enzyme cut DNA | ||
| Line 6: | Line 6: | ||
*Run a gel of linearized and cut DNA of BP1 and terminator assembly to verify transformation | *Run a gel of linearized and cut DNA of BP1 and terminator assembly to verify transformation | ||
*Begin ligation of standard RBS with BP1 gene to begin parallel assembly of our final operon | *Begin ligation of standard RBS with BP1 gene to begin parallel assembly of our final operon | ||
| - | |||
==Notes== | ==Notes== | ||
| - | *Maxiprepped | + | *Maxiprepped B0015 and B0034 smell strongly of alcohol, we are dubious of the content of the prepared DNA |
| + | ==Accomplished== | ||
| + | *Starting from maixprepped DNA we cut: | ||
| + | **E0240, I715039, B0034 (dubious, see above), BP1 | ||
| + | *Short Ligations: (45min) | ||
| + | **Test Vector 1: I715039 + E0240 | ||
| + | ***Did a ligation each for non-purified DNA (after digestion) and purified DNA. | ||
| + | **B0034 (RBS) + BP1 | ||
| + | ***Only non-purified post-digestion DNA | ||
| + | **Plated all short ligations | ||
| + | *Long Ligations: (overnight @ room temp) | ||
| + | **Test Vector 1: I715039 + E0240 (unpurified) | ||
| + | **B0034 (RBS) + BP1 | ||
| + | *Plated screening plasmid [http://partsregistry.org/Part:pSB1A10 pSB1A10] that we received today | ||
| - | [[Team:Virginia/Notebook|Notebook]] | + | [[Team:Virginia/Notebook|Back to Notebook]] |
Latest revision as of 22:19, 8 July 2008
Goals
- Prepare overnight broth of single successful colony of Promoter + RBS transformation
- Plate Screening plasmid (psb1a10) when it arrives from iGem
- Try again with ligation of RFP and GFP enzyme cut DNA
- Transform this ligation and pray that it grows in the incubator
- Run a gel of linearized and cut DNA of BP1 and terminator assembly to verify transformation
- Begin ligation of standard RBS with BP1 gene to begin parallel assembly of our final operon
Notes
- Maxiprepped B0015 and B0034 smell strongly of alcohol, we are dubious of the content of the prepared DNA
Accomplished
- Starting from maixprepped DNA we cut:
- E0240, I715039, B0034 (dubious, see above), BP1
- Short Ligations: (45min)
- Test Vector 1: I715039 + E0240
- Did a ligation each for non-purified DNA (after digestion) and purified DNA.
- B0034 (RBS) + BP1
- Only non-purified post-digestion DNA
- Plated all short ligations
- Test Vector 1: I715039 + E0240
- Long Ligations: (overnight @ room temp)
- Test Vector 1: I715039 + E0240 (unpurified)
- B0034 (RBS) + BP1
- Plated screening plasmid [http://partsregistry.org/Part:pSB1A10 pSB1A10] that we received today
