IGEM:IMPERIAL/2009/Assays Protocols/Cloning Strategy M1&T: Difference between revisions
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== Module 1 Cloning Strategy == | == Module 1 Cloning Strategy == | ||
[[Image:II09_clonstratm1.jpg]] | [[Image:II09_clonstratm1.jpg]] | ||
* Ligate PAH/ Cellulase gene sequence (G1) from GeneArt with the arabinose-inducible promoter from the iGEM starter kit. | * Ligate PAH/ Cellulase gene sequence (G1) from GeneArt with the arabinose-inducible promoter from the iGEM starter kit. | ||
* Test for successful ligation using | * Test for successful ligation using Cellulose/ PAH assay as defined [[IGEM:IMPERIAL/2009/Assays_Protocols | here]] | ||
* If successful, remove terminators from G1 gene sequence using suitable restriction enzymes. | * If successful, remove terminators from G1 gene sequence using suitable restriction enzymes. | ||
* Ligate sequence to LacI (BBa_C0012) from the iGEM starter kit. | * Ligate sequence to LacI (BBa_C0012) from the iGEM starter kit. | ||
* Test for expression of LacI gene using LacI repressible sequence coupled with reporter gene. BBa_I13601 is such a reporter - coupled with CFP. | |||
[[Image:II09_BBa_I13601.jpg]] | |||
<br> | |||
=== Preliminary Schematic === | |||
[[Image:II09_Clonstratm1_draft1.jpg]] | |||
Latest revision as of 09:04, 29 July 2009
Module 1 Cloning Strategy
- Ligate PAH/ Cellulase gene sequence (G1) from GeneArt with the arabinose-inducible promoter from the iGEM starter kit.
- Test for successful ligation using Cellulose/ PAH assay as defined here
- If successful, remove terminators from G1 gene sequence using suitable restriction enzymes.
- Ligate sequence to LacI (BBa_C0012) from the iGEM starter kit.
- Test for expression of LacI gene using LacI repressible sequence coupled with reporter gene. BBa_I13601 is such a reporter - coupled with CFP.
Preliminary Schematic
