This document is an instruction manual for HybriZAP-2.1 Two-Hybrid Libraries, providing information on the features and characteristics of the product.
The document serves as an instruction manual for the HybriZAP-2.1 Two-Hybrid Libraries system, which is designed for in vitro use to screen cDNA libraries. It details the contents of the kit, including premade libraries, control plasmids, bacterial host strains (XL1-Blue MRF' and XLOLR), yeast host strain (YRG-2), and helper phage.
The manual outlines comprehensive protocols for various stages of the two-hybrid system workflow. These include establishing bacterial stocks, determining library titer, preparing yeast host strains, storing and titering the helper phage, constructing DNA-binding domain vectors, performing yeast transformations (both simultaneous and sequential), and screening for interactions using filter lift assays and reporter gene expression verification.
| Library Type | Premade Library constructed in the HybriZAP-2.1 vector |
|---|---|
| Storage Temperature (Premade Libraries) | -80°C |
| Yeast Host Strain | YRG-2 |
| Helper Phage Titer | ~1.0 x 10^9 pfu/ml |
The premade library should be stored at -80°C upon arrival.
The library should not be passed through more than two freeze-thaw cycles.
The ExAssist interference-resistant helper phage has a titer of approximately 1.0 x 10^9 pfu/ml.
Control plasmids should be stored at -20°C.