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.

Streak plates of insert and vector from glycerol stocks

How to streak a plate

• Antibiotic Used:
• Incubation Time:
• Image of Plate after growth

Grow 5ml Cultures of each from a single colony

• Cellular chassis: ?
• Growth media: ?
• Antibiotic: ?
• Growth Temperature: ?C
• Spin speed: ?rpm
• Incubation Time: ?hrs
• Final OD₆₀₀:

Miniprep plasmid DNA from cultures

Prep concentration vector: ?ng/μl Prep concentration insert: ?ng/μl

Transform ligation product into competent cells

Can use electroporation or chemical transformation