Gateway cloning manual

To perform the BP clonase reaction, select the pDEST17 and pDONR – DTU PCR Product Entry clone sequences then go Cloning → Gateway Cloning. The Gateway cloning tool will identify the att sites present on the entry vector and Destination vector and confirm an LR reaction can be performed. In the test tube an LR reaction creates two new plasmid species. Gateway™ technology Gateway™ Technology is a universal cloning method that takes advantage of the site-specific recombination properties of bacteriophage lambda to provide a rapid and highly efficient way to move DNA sequences into multiple vector systems. The Gateway™ Technology is schematically represented www.doorway.ru Size: KB. directional TOPO cloning and direct access to the multitude of Gateway™ expression vectors. Figure 3. Strategies to build the Entry clone. The three possible methods that lead to the Entry clone are depicted: (A) BP cloning, (B) TOPO cloning, and (C) restriction enzyme and ligase cloning. Red arrows represent the fragment of www.doorway.ru Size: KB.

The Donor Vector box, the Gateway® BP Clonase® II Enzyme Mix and Reagents box, and the M13 Sequencing Primers box (Boxes 1–4) contain the following items. Store the Gateway® BP Clonase® II enzyme mix at −30°C to −10°C for up to 6 months. The Gateway cloning method, developed by Invitrogen, is an in vitro version of the integration and excision recombination reactions that take place when lambda phage infects bacteria. I n vivo, t hese recombination reactions are facilitated by the recombination of attachment sites from the phage (att P) and the bacteria (att B). directional TOPO cloning and direct access to the multitude of Gateway™ expression vectors. Figure 3. Strategies to build the Entry clone. The three possible methods that lead to the Entry clone are depicted: (A) BP cloning, (B) TOPO cloning, and (C) restriction enzyme and ligase cloning. Red arrows represent the fragment of interest.

This subcloning step is facilitated by the presence of a chlR gene within the Gateway cassette, allowing the desired clones to be selected based on their. 3 de out. de Creating Entry Clones Using the BP Recombination Reaction. This manual provides an overview of the MultiSite Gateway® Technology, and. Gateway Cloning Manual - Free download as PDF File .pdf), 5 Details of the GATEWAY Cloning Reactions6 Entry Vectors and Entry Clones.