BioBricks construction tutorial: Difference between revisions
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==Contacts== | ==Contacts== | ||
[[Barry Canton]], [[Jason Kelly]], [[Will Bosworth]] | [[Barry Canton]], [[Jason Kelly]], [[Will Bosworth]] | ||
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Assumed starting point is from glycerol stocks of insert and vector. | Assumed starting point is from glycerol stocks of insert and vector. | ||
== | ==[[Streak plates]] of insert and vector from glycerol stocks== | ||
== | ==[[Bacterial cell culture|Grow 5ml Cultures]] of each from a single colony== | ||
== | ==[[Miniprep]] plasmid DNA from cultures== | ||
== | ==[[Restriction digest]] insert and vector with appropriate enzymes== | ||
== | ==[[DNA Gel extraction|Extract]] vector and insert from gel== | ||
== | ==[[DNA Ligation|Ligate]] insert and vector== | ||
== | ==[[Electroporation|Transform]] ligation product into competent cells== | ||
Can use [[Electroporation|electroporation]] or [[Chemically competent cells|chemical transformation]] | Can use [[Electroporation|electroporation]] or [[Chemically competent cells|chemical transformation]] | ||
== | ==Screen colonies for correct plasmid by PCR== | ||
== | ==[[Miniprep]] DNA from chosen colony== | ||
== | ==Make glycerol and sequence DNA== | ||
Revision as of 15:17, 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
