AAV Quantification Kit
For the detection of adeno-associated virus (AAV) inverted terminal repeat (ITR) sequences in a real-time PCR based on the use of TaqMan® technology
AAV Quantification Kit
For the detection of adeno-associated virus (AAV) inverted terminal repeat (ITR) sequences in a real-time PCR based on the use of TaqMan® technology
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Features and Benefits
- Rapid AAV DNA extraction and quantification can be completed in 2 hours
- Universal qPCR assay targeting the AAV2 ITR enables quantification of most AAV vector constructs
- Purified DNA standard enables easy comparison of data between different lab groups
- Non-plasmid based standard prevents quantification artifacts due to insufficient melting of plasmid sequences
- DNAse digestion step aids in eliminating non-viral DNA
- Minimal sample handling for maximum AAV DNA recovery
Norgen’s AAV Quantification Kit is designed for the detection of adeno-associated virus (AAV) inverted terminal repeat (ITR) sequences in a real-time PCR based on the use of TaqMan® technology. A purified standard based solely on the AAV2 ITR sequence purified using Norgen’s proprietary silicon carbide technology simplifies the generation of a reliable standard curve for AAV quantification. Avoidance of a plasmid based standard eliminates problems associated with efficient melting of the ITR sequence due to coupling of the ITR to the much longer plasmid sequence, as well as variability due to rearrangements/duplications/deletions of the recombination prone ITR. An easy and rapid method for viral DNA extraction simplifies the step of obtaining AAV DNA while simultaneously eliminating contaminating non-encapsidated DNA. Norgen’s AAV Quantification Kit can facilitate pre-clinical studies that require accurate vector titration as well as interlab comparisons of vector quantities. This kit is designed for research use only and not for use in diagnostic procedures.
Details
Supporting Data
Storage Conditions and Product Stability
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots. All reagents can be stored for 1 year at -20°C without showing any reduction in performance.
Component | Cat. 63800 (24 rxns) |
---|---|
Enzyme Incubation Buffer A | 500 µL |
DNase I | 25 µL |
DNase Stop Solution | 100 µL |
2X PCR Mastermix | 350 µL |
AAV Primer & Probe Mix | 90 µL |
AAV Standard | 6 µL |
Nuclease-Free Water (Negative Control) | 1.25 mL |
Product Insert | 1 |
Documentation
FAQs
The AAV quantification is a universal qPCR assay targeting the AAV2 ITR, and enables quantification of most AAV vectors. The AAV2 ITR Quant Kit has a cross reactivity for AAV1, 3, 6, and 7.
This kit can detect from approx. 2 x 10^3 to 2 x 10^10 genome copies of AAV.
This kit uses a 62 bp amplicon covering the AAV2 ITR to overcome the limitations of using a plasmid-based standard.
Citations
Title | Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screening |
Journal | Scientific Reports.2022. |
Authors | Lea Jessica Flitsch, Kathleen Börner, Christian Stüllein, Simon Ziegler, Vera Sonntag-Buck, Ellen Wiedtke, Vesselina Semkova, Si Wah Christina Au Yeung, Julia Schlee, Mohamad Hajo, Mona Mathews, Beatrice Stefanie Ludwig, Susanne Kossatz, Horst Kessler, Dirk Grimm & Oliver Brüstle |