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Team:Cambridge/Notebook/Week4
From 2010.igem.org
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# Digest luciferase Biobrick with: XbaI, PstI | # Digest luciferase Biobrick with: XbaI, PstI | ||
# Digest backbone Biobrick with SpeI, PstI | # Digest backbone Biobrick with SpeI, PstI | ||
| - | # Run on | + | # Run both digest products on a gel |
# Extract suitable lengthed fragments from gel: | # Extract suitable lengthed fragments from gel: | ||
## Backbone should be 2153 bp | ## Backbone should be 2153 bp | ||
Revision as of 18:12, 4 August 2010
Notebook: Week 4
Monday
- Wet work
- Transformed competent cells using plasmid phK724 from Jim Slock (thanks!)
- Grew up E. coli containing phK555 in 5ml LB broth in rotating 37°C incubator overnight
- Dry work
- Finalising sequencing requirements
- Dismissed PPK
- Decided to include Luciola cruciata
- Further Wiki work
- Finalising sequencing requirements
Tuesday
- Wet Work
- Made competent cells of 4 different hns mutants: W3110 hns93-1, BW25113 Δhns::kan, W3110 hns-205::Tn10, GM230 hns-205::Tn10
- Now stored in the -80°C freezer for later use
- Dry Work
- Organising sequences for synthesis
Wednesdy
Thursday
Theo's plan
- Miniprep plasmid DNA from luciferase and backbone transformed TOP10 following Qiagen protocol
- Digest luciferase Biobrick with: XbaI, PstI
- Digest backbone Biobrick with SpeI, PstI
- Run both digest products on a gel
- Extract suitable lengthed fragments from gel:
- Backbone should be 2153 bp
- Luciferase should be 1653 bp (its 3426 bp backbone should be discarded)
- Perform ligation
- Transform competent TOP10 cells with ligation product
Friday
Saturday
Sunday
