BioBricks construction tutorial

Contacts

Barry Canton, Jason Kelly, Will Bosworth

Aim

Describe the steps of a standard BioBricks assembly for two parts and include images of expected results (e.g., gels, seq files, etc). This will provide a walkthrough for someone new to BioBricks construction to practice with parts that are known together well.

Steps

Assumed starting point is from glycerol stocks of insert and vector.

1. Streak plates of Insert and Vector from glycerol stocks.
2. Grow 5ml Cultures of each from a Single Colony
3. Miniprep Plasmid DNA from Cultures
4. Restriction digest Insert and Vector with Appropriate Enzymes
5. Extract Vector and Insert from Gel
6. Ligate Insert and Vector
7. Transform Ligation Product into Competent Cells
   • Can use electroporation or chemical transformation
8. Screen Colonies for Correct Plasmid by
9. Miniprep DNA from Chosen Colony
10. Make Glycerol and Sequence DNA