Team:Cambridge/Notebook/Week4

(Difference between revisions)

Revision as of 18:06, 4 August 2010

Notebook: Week 4

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

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

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 on gels
Extract suitable lengthed fragments from gel:
1. Backbone should be 2153 bp
2. Luciferase should be 1653 bp (its 3426 bp backbone should be discarded)
Perform ligation
Transform competent TOP10 cells with ligation product

@@ Line 23: / Line 23: @@
 ==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 on gels
+# 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==