BioBricks construction tutorial: Difference between revisions
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==[[Miniprep]] plasmid DNA from cultures== | ==[[Miniprep]] plasmid DNA from cultures== | ||
Prep concentration vector: ?ng/μl | *Prep concentration vector: ?ng/μl (?μl elution volume) | ||
Prep concentration insert: ?ng/μl | *Prep concentration insert: ?ng/μl (?μl elution volume) | ||
==[[Restriction digest]] insert and vector with appropriate enzymes== | ==[[Restriction digest]] insert and vector with appropriate enzymes== |
Revision as of 15:28, 25 August 2005
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
- 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 (?μl elution volume)
- Prep concentration insert: ?ng/μl (?μl elution volume)
Restriction digest insert and vector with appropriate enzymes
Extract vector and insert from gel
Ligate insert and vector
Transform ligation product into competent cells
Can use electroporation or chemical transformation