AAV Quantification Kit

Overview

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

Figure 1 / 3

Figure 1. Amplification of ten fold serial dilutions of purified Norgen AAV Standard: 10-2 to 10-8 dilutions and no target control (NTC).

Figure 2. XY Scatter Plot of Ct versus log copy number demonstrates high linearity for AAV standard dilution series from 10-2 dilution to 10-8 dilution (copy number of 2,000 to 2,000,000,000).

Figure 3. A) Determination of copy number using Norgen's AAV Quantification Kit for samples purified with Norgen's AAV Purification Kit based on 1 mL, 3 mL, or 9 mL input volumes. B) Transduction of HTX cells with 100 µL of AAV preparation from 1 mL, 3 mL, or 9 mL input volumes purified using the Norgen AAV Purification Kit.

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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

Component	Cat. TM30850 (100 preps)	Cat. TM30810 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
S. uberis Primer & Probe Mix	280 μL	280 μL
S. uberis Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

Detail

Overview

Details

Supporting Data

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