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
Assembly reactions (BsaI/BsmBI)
Golden Gate Assembly is a
molecular cloning method used to recombine multiple DNA components into a single linear piece or plasmid. It bears similarity to
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
*
--
Victor Li - 2021-11-03
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