TesA/RBS Ligation Attempt 2
- Checked DNA concentrations
- TesA= 11.99 ng/μL ; RBS=32.99 ng/μL
- When divided by the number of base pairs: [TesA]=.018 and [RBS]=.016
| Reagent (μL)
|| Volume (μL)
| DNA(plasmid) || 1.0 μL RBS, 2.0 μL TesA
| 10X buffer || 1.0
| dH2O || 5.0
| Ligaze || 1.0
| || 10.0 μL --> 25°C/ ~1 Hr.
- After ligation, the mixture was heated to 65°C for 10 min. to denature the ligaze.
- Transformation was then performed.
- Three plates were grown overnight: RBS backbone (vector), TesA+RBS, and mCherryboth as a positive control and to replenish our stocks.
- Unfortunately, no growth was seen on any of the plates, including the positive control of mCherry.