Agilent pDual Expression Vector Instruction Manual Catalog #214501 and #214502

About this manual

The document is a user instruction manual for the Agilent pDual Expression Vector, available in two catalog sizes (#214501 containing 20 μg and #214502 containing 100 μg). The vector is designed for constitutive expression in mammalian cells via the CMV promoter and inducible gene expression in prokaryotes via a hybrid T7/lacO promoter.

The manual details the materials provided, including the vector plasmid and XL1-Blue host strain. It outlines protocols for cloning inserts using PCR amplification with specific primers containing Eam1104 I recognition sites to create directional seamless junctions. The guide covers digesting the vector, preparing the insert, ligating the components, and transforming or transfecting into appropriate bacterial or mammalian cells.

What's inside

• Materials Provided
• Storage Conditions
• Introduction
• Insert Primer Design
• Digesting pDual Expression Vector
• Preparing PCR Amplified Insert
• Ligating Vector and Insert
• Transforming/Transfecting Ligated Vector/Insert

Specifications

Frequently asked questions

What is the recommended incubation temperature for digesting the pDual expression vector?

The reaction should be incubated at 37°C for 2 hours.

Which restriction enzyme is recommended for digesting the pDual expression vector?

The Eam1104 I restriction endonuclease is recommended to generate a ligation-ready vector.

What type of cells should be used for the initial transformation?

An endA– strain such as XL1-Blue supercompetent cells is recommended for the initial transformation.