Team:Wash U/Protocol
From 2009.igem.org
(Difference between revisions)
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+ | =='''Mini-Prep'''== | ||
+ | <font size="2"> | ||
+ | :general description | ||
+ | '''Materials''' | ||
+ | :List of materials | ||
+ | '''Procedures''' | ||
+ | :List of Procedures | ||
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- | + | =='''Biobrick Assembly'''== | |
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- | ''' | + | :General description |
+ | '''Materials''' | ||
+ | :List of materials | ||
+ | '''Procedures''' | ||
:1. Begin BioBrick Assembly with three separate plasmids, an upstream part, a down stream part, and a destination plasmid. It is important that the destination plasmid contain the toxic gene ccdB in the BioBrick cloning site and a different antibiotic resistance to the upstream and downstream parts.<br> | :1. Begin BioBrick Assembly with three separate plasmids, an upstream part, a down stream part, and a destination plasmid. It is important that the destination plasmid contain the toxic gene ccdB in the BioBrick cloning site and a different antibiotic resistance to the upstream and downstream parts.<br> | ||
:2. Digest each part with the proper restriction enzymes to isolate the upstream and downstream parts, and to remove the BioBrick from the destination plasmid.<br> | :2. Digest each part with the proper restriction enzymes to isolate the upstream and downstream parts, and to remove the BioBrick from the destination plasmid.<br> | ||
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- | '''Digestion''' | + | =='''Digestion'''== |
+ | <font size="2"> | ||
+ | :General Description | ||
+ | '''Materials''' | ||
+ | :list of materials | ||
+ | '''Procedures''' | ||
:Note: The procedures from this point forward assume that DNA has been amplified and purified. If this is not the case, please read the Polymerase Chain Reaction procedure for amplification and the Mini-Prep procedure for purification.<br> | :Note: The procedures from this point forward assume that DNA has been amplified and purified. If this is not the case, please read the Polymerase Chain Reaction procedure for amplification and the Mini-Prep procedure for purification.<br> | ||
:1. Begin by thawing the upstream, downstream, and destination plasmid parts along with the NEBuffer 2 and BSA.<br> | :1. Begin by thawing the upstream, downstream, and destination plasmid parts along with the NEBuffer 2 and BSA.<br> | ||
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- | '''Ligation''' | + | =='''Ligation'''== |
+ | <font size="2"> | ||
+ | :General description | ||
+ | '''Materials''' | ||
+ | :List of Materials | ||
+ | '''Procedures''' | ||
:Note: this procedure requires the products of a successful digestion.<br> | :Note: this procedure requires the products of a successful digestion.<br> | ||
:1. Thaw the 10X T4 DNA Ligase Reaction Buffer and mix to dissolve the precipitate.<br> | :1. Thaw the 10X T4 DNA Ligase Reaction Buffer and mix to dissolve the precipitate.<br> | ||
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- | '''Gel Electrophoresis''' | + | <font size="4"> |
+ | =='''Gel Electrophoresis'''== | ||
+ | <font size="2"> | ||
+ | :General Description | ||
+ | '''Materials''' | ||
+ | :List of Materials | ||
+ | '''Procedures''' | ||
:text | :text | ||
Revision as of 16:04, 6 July 2009