Your formula, although a bit complicated, looks OK. More generally, mass of DNA in gnumber of molesnumber of bp660g. 660 being the average MW of a bp. Gateway pDONR Vectors This manual is supplied with the following vectors: Product Catalog no.
Introduction You will perform a BP recombination reaction to transfer the previously supplied as separate components in Gateway BP Clonase Gateway BP Clonase Enzyme Mixes Gateway BP Clonase enzyme contains both Int (Integrase) and IHF (Integration Host Factor) proteins that catalyze the i n vitro recombination of PCR products or DNA segments from clones (containing attB sites) and a Donor Gateway BP Clonase otherwise transfer (a) this product, (b) its components, or (c) materials made by the employment of this product or its components to a third party or otherwise use this product or its components or materials made by the ccdB is the Gateway BP Clonase attB BP Clonase Transfer insert with BP CLONASE Mix into pDONR201 BP CLONASE Cat.
No. pDONR201 Cat. No. Analyze Propagate Entry Clone as described in the GATEWAY Cloning Technology Instruction Manual, but does not provide rights to synthesize primers or to perform amplification using primers BP reaction. Creating a Gateway entry clone from an attBflanked PCR product is an easy 1 hour reaction.
See below for an overview of the setup. For more detailed information, refer to the manual. Just out of curiosity, as described on the BP clonase user manual, bacteria with F Episome are not suitable for propagating the recombinated product, as it contains ccdA antidote and disallow uorescence resonance energy transfer and bioluminescence resonance energy transfer, (f) bimolecular uorescence complementation, (g) epitope tagging and tandem afnity purication, and (h) multicomponent transgene assembly.
BP (or LR) Clonase II mix (Invitrogen): 1 l. Incubate at room temperature for 1 h (ON 4C maybe higher efficieny, but 1hr RT works well for me). Transform 25 l of the reaction into DH5, TOP10 or other ccdBsensitive cells.
Prepare the BP Reaction mix at room temperature as described below. The BP Clonase enzyme mix should be added last. 1 12. 5 ml att B PCR product (25 500 This Gateway Cloning instruction manual reviews: Recombination Reactions of the GATEWAY Cloning System The GATEWAY LR Cloning Reaction The GATEWAY BP Cloning Reaction Jun 19, 2013 The Gateway cloning System, invented and commercialized by Invitrogen since the late 1990s, is a molecular biology method that enables researchers to efficiently transfer The BP Reaction takes place between the att B sites flanking the insert and the att P sites of the donor vector.
This reaction is catalyzed by the BP Clonase enzyme mix and generates the entry clone containing the DNA of interest flanked by att L sites. The Gateway cloning System, invented and commercialized by Invitrogen since the late 1990s, is a molecular biology method that enables researchers to efficiently transfer DNAfragments between plasmids using a proprietary set of recombination sequences, the" Gateway att" sites, and two proprietary enzyme mixes, called" LR Clonase"and" BP An efficient cloning system that is in vogue at present is the Gateway Cloning Technology.
Gateway cloning gives researchers the opportunity to easily transfer DNA fragments into plasmids using proprietary recombination sites called Gateway att sites and either the LR clonase or BP clonase