WebThe two steps of Golden Gate assembly. (A) Individual parts are provided as plasmid-borne cargo or linear PCR products and mixed in a single tube together with the restriction enzyme (scissors) and the DNA ligase … WebGolden Gate assembly reagents Adenosine triuphosphate (ATP) (10 mM) (e.g., New England BioLabs) Bovine serum albumin (BSA) (10×) (e.g., New England BioLabs) NEBuffer 4 (New England BioLabs) NEB CutSmart buffer can be used in place of NEBuffer 4 and BSA. pBH-donor vector DNA (available from authors on request)
Golden Gate Assembly NEB
WebAvoid PCR-induced errors for amplicon inserts/modules. Do not over-cycle and use a proofreading high fidelity DNA polymerase, such as Q5 ® DNA High-Fidelity Polymerase. … WebMar 8, 2024 · Assembly efficiency was thus greater than that of my conventional Golden Gate assembly and background was lower, thereby validating the design of the 5RM workflow. The vector only control, however, produced a surprisingly high 147 ± 43 colonies, which suggested that the Prham - M.EcoRI - M.XbaI - M.SalI -p15A- aadA expression … classlink problems
The Golden Gate assembly method (and MoClo and …
WebAn E. coli strain that carries the cloned PluTI gene from Photorhabdus luminescens This product is related to the following categories: Restriction Endonucleases P R This product can be used in the following applications: Golden Gate Assembly Reagents Supplied Reagents Supplied The following reagents are supplied with this product: WebJul 13, 2024 · Golden Gate is a molecular cloning method that facilitates the assembly of multiple DNA fragments into a single piece. This method relies on the presence of restriction sites within a particular sequence to be cloned. It originated in 1996. This technique uses Type IIS restriction enzymes and T4 DNA ligase. WebThe NEBridge Golden Gate Assembly Kit (BsmBI-v2) contains an optimized mix of BsmBI-v2 and T4 DNA Ligase. BsmBI-v2 has been engineered by NEB and outperforms BsmBI in Golden Gate Assemblies. Together these enzymes can direct the assembly of multiple inserts/modules and also single insert/library generation cloning with single insert (s) … classlink rcstn launchpad