(r3) GoldenGateAssembly < Lab

---+ 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%

Barrick Lab > ProtocolList > GoldenGateAssemblyProtocolsMainPage > GoldenGateAssembly

Topic revision: r3 - 2023-02-08 - 16:39:45 - Main.CameronRoots