BioBricks construction tutorial: Difference between revisions
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#[[Bacterial cell culture|Grow 5ml Cultures]] of each from a Single Colony | #[[Bacterial cell culture|Grow 5ml Cultures]] of each from a Single Colony | ||
#[[Miniprep]] Plasmid DNA from Cultures | #[[Miniprep]] Plasmid DNA from Cultures | ||
#[[ | #[[Restriction digest]] Insert and Vector with Appropriate Enzymes | ||
#[[DNA Gel extraction|Extract]] Vector and Insert from Gel | #[[DNA Gel extraction|Extract]] Vector and Insert from Gel | ||
#[[DNA Ligation|Ligate]] Insert and Vector | #[[DNA Ligation|Ligate]] Insert and Vector | ||
Revision as of 14:16, 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.
- Grow 5ml Cultures of each from a Single Colony
- Miniprep Plasmid DNA from Cultures
- 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
- Screen Colonies for Correct Plasmid by
- Miniprep DNA from Chosen Colony
- Make Glycerol and Sequence DNA
