From 2011.igem.org
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Revision as of 16:55, 4 October 2011
| Lab Diaries
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| Week 1
Transformation of reporter DNA (pEGFP-loxp-km-loxp) into DH10B (non-virulent strain E. coli) with antibiotic resistance (Chloramphenicol – Cm)
| Lab Diaries
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Week 2
- tetO2 -1 (DNA binding site)
- Reverse PCR was used to insert tetO2 -1 into pEGFP-loxp-km-loxp (template DNA) by using forward and reverse primers which are with the tetO2 -1. By using reverse PCR, we can insert the tetO2 -1 site into the pEGFP while the plasmid produced is still in double-stranded circular form.
- pEGFP-loxp-km-loxp-tetO2-1 was then transformed into DH10B to greatly amplify the product by using bacterial cells
- tetO2 -2 (DNA biding site)
- Reverse PCR was used to insert tetO2 -2 into pEGFP-loxp-km-loxp
- 2uL of 10-fold diluted pEGFP-loxp-km-loxp- tetO2 -2 was transformed into DH10B to greatly amplify the product by using bacterial cells
- tetO2 – 3 (DNA binding site)
- Reverse PCR was used to insert tetO2 – 3 into pEGFP-loxp-km-loxp
- 2uL of 10-fold diluted pEGFP-loxp-km-loxp- tetO2 – 3 was transformed into DH10B to greatly amplify the product by using bacterial cells
| Lab Diaries
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Week 3
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| Fusion protein genes produced separately using PCR
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