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Team:Paris/Parts/Plastest
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| - | + | = Plasmid for promoter Amplification & Measurement = | |
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Instead of using a big biobrick containing Promoter, RBS, GFP and Terminators, the Biobrick used for the test is only the Promoter. | Instead of using a big biobrick containing Promoter, RBS, GFP and Terminators, the Biobrick used for the test is only the Promoter. | ||
This plasmid can be used to amplify a promoter because only the promoter is in the biobrick. | This plasmid can be used to amplify a promoter because only the promoter is in the biobrick. | ||
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| - | + | One derivated plasmid do not have the RBS B0032 outside the BioBrick site, it allows to test the influence of the RBS in changing it in the BioBrick. To do so, [[team:Paris/Notebook/Oligo|O142]] does not contains the RBS, the PCR will begin directly at the ATG codon. | |
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| + | == Oligonucleotides design == | ||
Creation of 3 oligonucleotides ([[team:Paris/Notebook/Oligo|O140, O141, O142]]) in order to create two particular plasmids. | Creation of 3 oligonucleotides ([[team:Paris/Notebook/Oligo|O140, O141, O142]]) in order to create two particular plasmids. | ||
[[image:oligo.jpg|frame|center|Design of the oligonucleotides [[team:Paris/Notebook/Oligo|O140 and O141]] ]] | [[image:oligo.jpg|frame|center|Design of the oligonucleotides [[team:Paris/Notebook/Oligo|O140 and O141]] ]] | ||
| - | == | + | == Construction of the Plasmid == |
| - | + | Necessary material | |
| - | * Oligos | + | * Oligos O140, O141 and O142 |
| - | + | * E0240 | |
| - | + | * pSB3K3 | |
Latest revision as of 06:50, 30 October 2008
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Plasmid for promoter Amplification & Measurement
Those plasmids are very useful vectors to amplify promoters and to measure their forces using Standard Promoter Units. The principle is easy to understand. The plasmid contains the BioBrick restriction sites (EcoRI, XbaI, SpeI and PstI) followed by the standard GFP Tripart (E0240). Instead of using a big biobrick containing Promoter, RBS, GFP and Terminators, the Biobrick used for the test is only the Promoter. This plasmid can be used to amplify a promoter because only the promoter is in the biobrick.
One derivated plasmid do not have the RBS B0032 outside the BioBrick site, it allows to test the influence of the RBS in changing it in the BioBrick. To do so, O142 does not contains the RBS, the PCR will begin directly at the ATG codon.
Oligonucleotides design
Creation of 3 oligonucleotides (O140, O141, O142) in order to create two particular plasmids.
Construction of the Plasmid
Necessary material
- Oligos O140, O141 and O142
- E0240
- pSB3K3
