Team:Wash U/Protocol
From 2009.igem.org
(Difference between revisions)
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=='''Digestion'''== | =='''Digestion'''== | ||
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- | : | + | :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. |
'''Materials''' | '''Materials''' | ||
:list of materials | :list of materials | ||
'''Procedures''' | '''Procedures''' | ||
- | |||
# Begin by thawing the upstream, downstream, and destination plasmid parts along with the NEBuffer 2 and BSA. | # Begin by thawing the upstream, downstream, and destination plasmid parts along with the NEBuffer 2 and BSA. | ||
# In three separate PCR microcentrifuge tubes labeled upstream, downstream, and destination, add 500ng of the respective dried DNA and dilute with dH20 to 42.5 uL. | # In three separate PCR microcentrifuge tubes labeled upstream, downstream, and destination, add 500ng of the respective dried DNA and dilute with dH20 to 42.5 uL. | ||
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=='''Ligation'''== | =='''Ligation'''== | ||
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- | Ligation formally joins two or more pieces of DNA together that are already annealed. Note: this procedure requires the products of a successful digestion. | + | :Ligation formally joins two or more pieces of DNA together that are already annealed. Note: this procedure requires the products of a successful digestion. |
- | + | ||
'''Materials''' | '''Materials''' | ||
:List of Materials | :List of Materials |
Revision as of 20:11, 8 July 2009