---+ Golden Gate Assembly *Under Construction* Refer to Bee Toolkit pages for design and assembly of parts and plasmids for use with Bee Toolkit [[Generating Overhangs][Generating Overhangs]] [[Assembly reactions][Assembly reactions (BsaI/BsmBI)]] Golden Gate Assembly is a [[https://en.wikipedia.org/wiki/Molecular_cloning][molecular cloning]] method used to recombine multiple DNA components into a single linear piece or plasmid. It bears similarity to [[https://blog.addgene.org/plasmids-101-restriction-cloning][restriction cloning]] by using a restriction endonuclease to produce "sticky ends", allowing for two terminal ends of DNA (either two distinct DNA molecules or a single DNA linear DNA molecule circularizing) to hybridize and be annealed together via a DNA Ligase. To perform Golden Gate Assembly, one or more Type IIS Restriction Enzymes are used. Commonly used enzymes in the Barrick Lab are: * BsmbI * BsaI * PaqCI ----+ Reagents * Your Restriction Enzyme(s) of choice * T4 ligase Buffer * -- %USERSIG{VictorLi - 2021-11-03}% ---++ Comments %COMMENT%
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Topic revision: r3 - 2023-02-08 - CameronRoots