6x50ml Angle Rotor 

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

Description 

The PM2610/PM2611 ExcelBand™ Enhanced 3-color High Range Protein Marker is a ready to use three-color protein standard with 12 pre-stained proteins covering a wide range of molecular weights from 10 to 245 kDa in Tris-Glycine buffer (9 to 235 kDa in BisTris (MOPS) buffer and 10-235 kDa in Bis-Tris (MES) buffer). Proteins are covalently coupled with different chromophores for easy identification of bands, with two reference proteins carrying enhanced intensity corresponding to a green at 25 kDa and red at 75 kDa, respectively, as separated on SDS-PAGE (Tris-Glycine buffer). The ExcelBand™ Enhanced 3-color High Range Protein Marker is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins.

Features

• Ready-to-use — Premixed with a loading buffer for direct loading, no need to boil.
• Two reference bands — 75 kDa (red) and 25 kDa (green)

Contents 

Approximately 0.2~0.6 mg/ml of each protein in the buffer (20 mM Tris-phosphate (pH 7.5 at 25°C), 2 % SDS, 3.6 M Urea, and 15 % (v/v) Glycerol). 

Quality Control 

Under suggested conditions, the PM2610/PM2611 Enhanced 3-color High Range Protein Marker resolves 12 major bands in 15% SDS-PAGE (Tris-Glycine buffer, MOPS, and MES buffer) and after Western blotting to nitrocellulose membrane. 

Storage

4°C for 3 months 
-20°C for long term storage

The PM2610/PM2611 ExcelBand™ Enhanced 3-color High Range Protein Marker is a ready to use three-color protein standard with 12 pre-stained proteins covering a wide range of molecular weights from 10 to 245 kDa in Tris-Glycine buffer (9 to 235 kDa in BisTris (MOPS) buffer and 10-235 kDa in Bis-Tris (MES) buffer). Proteins are covalently coupled with different chromophores for easy identification of bands, with two reference proteins carrying enhanced intensity corresponding to a green at 25 kDa and red at 75 kDa, respectively, as separated on SDS-PAGE (Tris-Glycine buffer). The ExcelBand™ Enhanced 3-color High Range Protein Marker is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins.