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Team:Hawaii/Antibiotic test for aadA construct
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Contents |
Antibiotic test for aadA construct
- The gene for Sp and Sm resistance was constructed in BioBrick format. It was amplified from pRL1383a with primers adding BioBrick ends, and the lac promoter and rbs were ligated to it.
- This construct was tested for antibiotic resistance.
Methods
- Sequencing confirmation
- Streak on LB+Agar plates with Sm50Sp10
- constructs
- (+) control pSMC121 (contains aadA gene)
- (-) control pSB1A3
- Transformation of the plasmid from one of these constructs into E. coli and plate on LB+Agar with varying concentrations of SmSp.
- DH5-a
- DB3.1
- controls: (+) pSMC121, (-) no plasmid
Results
- Sequencing confirmation
- Streak
The plate streaked with multiple aadA constructs + controls.
- All four of the constructs grew and the positive control grew while the negative control did not grow.
Discussion
- What was learned and how to do future experiments differently.
