Golden Gate Assembly

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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