This document provides instructions for using the AAV Helper-Free System and information on the limited product warranty. The product is for research use only and not for diagnostic procedures.
The document serves as an instruction manual for the Agilent AAV Helper-Free System (Catalog #240071) and associated control plasmids. It outlines the contents provided with the kit, including specific vectors like pAAV-MCS and pHelper plasmid, along with cell lines such as AAV-293 and AAV-HT1080.
The guide covers essential laboratory procedures for researchers, starting with safety considerations regarding Biosafety Level 2+ protocols. It details the generation of recombinant adeno-associated virus (AAV) stocks through transfection of AAV-293 cells, monitoring virus production, and preparing viral stocks. Additionally, it includes guidelines for titering using AAV-HT1080 cells and methods for verifying cloning results.
| pAAV-MCS vector concentration | 1 μg/μl in TE buffer |
|---|---|
| pHelper plasmid quantity | 20 μg (sufficient for two transfections) |
| AAV-293 cells quantity | 1 × 106 cells |
| Storage temperature for AAV Vectors | -20°C |
Production of adeno-associated virus and use of AAV vectors fall within NIH Biosafety Level 2 criteria.
The manual recommends using competent E. coli cells such as XL10-Gold ultracompetent cells.
The system includes 20 μg of pHelper plasmid at a concentration of 1 μg/μl.