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

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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

Introduction

MagPure Stool DNA Kit is specially designed for high throughput DNA extraction from stool samples. It can get high purity microbial DNA from stool samples (≤200mg). This kit is based on magnetic beads purification and unique inhibiting factors adsorption technology, no phenol-chloroform extraction or alcohol precipitation. It can adsorb humic acid and other inhibiting factors in the solution efficiently. DNA can be directly used for downstream applications such as PCR, Viral DNA testing, bacterial DNA testing, etc.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 100-150mg stool samples
Applications	PCR, southern blot and enzyme digestion, etc.
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Stool
Sample amount	100-150 mg
Elution volume
Time per run

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Lysozyme. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• Fast – several samples can be extracted in 60 minutes (after digestion)
• High purity – unique adsorbent can completely remove inhibitory factors
• High recovery – DNA can be recovered at the level of PG

Kit Contents

Contents	D6364
Purification Times	400 Preps
MagPure Particles N	14 ml
2ml Bead Tubes	400
PVP-10	6 g
RNase A	75 mg
Proteinase K	180 mg
Protease Dissolve Buffer	20 ml
Buffer ATL	300 ml
Buffer PCI	300 ml
Buffer MLE	180 ml
Buffer GW1*	132 ml
Elution Buffer	60 ml

Storage and Stability

MagPure Particles, RNase A and Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

MagPure Stool DNA Kit is specially designed for high throughput DNA extraction from stool samples. It can get high purity microbial DNA from stool samples (≤200mg). This kit is based on magnetic beads purification and unique inhibiting factors adsorption technology, no phenol-chloroform extraction or alcohol precipitation. It can adsorb humic acid and other inhibiting factors in the solution efficiently. DNA can be directly used for downstream applications such as PCR, Viral DNA testing, bacterial DNA testing, etc.

Description

Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.

The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.

Packaged, optimised and ready to use. Expect Better Data.

Primer and probe mix (150 reactions)
Reverse Transcription, target specific primers (RNA genome viruses only)
Copy number standard curve (sufficient for multiple standard curves)
Internal extraction control – Read through VIC channel*
Endogenous control (150 tests)
RNAse/DNAse free water
*alternative fluorophores available on request