Reagents & Protocols

Alpha-based B1H binding site selection system:

The Strains and plasmids associated with our 2005 Nature Biotechnology paper are now available through addgene at www.addgene.org/Scot_Wolfe.

Additional useful files are provided below:

Overview of components from addgene

Annotations for plasmids

pH3U3-pB1H1-mcs.tif

pB1H1.asc pB1H1.tif

pB1H2.asc pB1H2.tif

pH3U3-mcs.asc pH3U3.tif

pB1H1-zif268.asc

pH3U3-Zif268.asc

Electrocomp cell prep instruct

NM medium & AA mix

Omega-based B1H binding site selection system components described in 2008 in Cell and NAR are available through Addgene.

Reagent list necessary to use the omega-based B1H system for the selection of Zinc Finger Proteins for the creation of ZFNs for targeted gene knockouts as described in our recent paper in Nature Biotechnology. A basic outline for testing the B1H reagents in your hands to confirm that they work before you get started.

We also have developed a Web-based ZFN target sequence analysis tool for identifying ZFN targets in genes and recommending the libraries and oligos that need to be used. A Protocol for ZFP library construction in omega B1H system provides more detailed infomation on the construction of a randomized library using our system.

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