This document provides instructions for the use of BL21-Gold Competent Cells, BL21-Gold(DE3) Competent Cells, and BL21-Gold(DE3)pLysS Competent Cells, as well as information on technical services. These cell lines are intended for research use only and are not for use in diagnostic procedures. The document also includes ordering information for the products.
The document is an instruction manual for three specific strains of E. coli competent cells: BL21-Gold, BL21-Gold(DE3), and BL21-Gold(DE3)pLysS. These high-performance cells are designed for protein expression studies utilizing the T7 RNA polymerase promoter. They feature improved transformation efficiency (greater than 1 x 10^8 cfu/ug of pUC18 DNA) and lack specific proteases like Lon and OmpT to prevent recombinant protein degradation.
The manual details the materials provided with each catalog number, storage conditions requiring -80°C, and specific transformation guidelines. It outlines protocols for transforming cells using heat pulses, inducing target proteins via IPTG or infection with Lambda CE6 phage, and growing bacteriophage stocks. Additionally, it includes troubleshooting tips, media preparation instructions, and an academic assurance letter regarding the use of T7 technology.
| Storage Temperature | -80°C |
|---|
The competent cells must be stored at -80°C and should not be kept in liquid nitrogen.
All three BL21-Gold-derived expression strains are resistant to tetracycline, and the BL21-Gold(DE3)pLysS strain is also resistant to chloramphenicol.
These strains naturally lack the Lon protease and are engineered to be deficient for the OmpT protein, both of which can degrade recombinant proteins.