CGT Viral Vector Lysis kit (100 rxn)

QIAGEN's CGT Viral Vector Lysis Kit is a sample-processing workflow for the determination of viral vector genome titers, particularly from Adeno-Associated Virus (AAV) particles. The kit supports a four-step process: DNase I treatment to remove DNA impurities such as residual host cell DNA or plasmids, capsid lysis (achievable without the need for Proteinase K, although a Proteinase K-based protocol is also described), serial dilution of the raw lysate, and quantification via digital PCR.

It is suitable for the lysis of AAV2, AAV5, AAV6, AAV8 and AAV9, and the resulting lysates are optimized for downstream use with QIAcuity Cell and Gene Therapy dPCR Assays in combination with the QIAcuity Probe PCR Kit on the QIAcuity dPCR System, delivering absolute quantification of encapsidated genome copies. Assays are provided as a ready-to-use 20× primer–probe mix in three fluorophore choices (FAM, HEX, and Cy5) for singleplex and multiplex titer determination. The kit is shipped in two boxes: Box 1 at ambient temperature (stored at 15–25°C), with DNase I vials held at 2–8°C, and Box 2 shipped on dry ice and stored at −30 to −15°C. Each lot is tested against predetermined specifications under QIAGEN's ISO-certified Quality Management System. The products are intended for molecular biology applications and are not intended for the diagnosis, prevention, or treatment of a disease.

In a published application, two AAV2 samples were processed with the kit and quantified on the QIAcuity Digital PCR instrument with 8.5K Nanoplates using the CGT dPCR Assays run in triplex reactions in the FAM, HEX and Cy5 channels. Samples were serially diluted in 6 steps from 15,000 copies/µL down to 2.5 copies/µL with an R²=1.0 on 8.5K Nanoplates, with each dilution measured in technical triplicates.