t-Boc-N-Amido-PEG2-propargyl

Description 

The G-HiFi™ DNA Polymerase is a new genetically modified, recombinant DNA polymerase suitable for GC-rich templates that are difficult to amplify. The fidelity of G-HiFi™ DNA Polymerase is 70 times higher than that of Taq DNA polymerase. The high extension rate of G-HiFi™ DNA Polymerase is achieved by blending the DNA polymerase with an elongation enhancer. The optimized 5X G-HiFi™ Buffer includes special ingredients that suppress non-specific amplification as well as plateau effect produced by conventional PCR. With the optimized 5X G-HiFi™ Buffer, G-HiFi™ DNA Polymerase is capable to amplify most templates, such as longer targets (up to 40 kb from lambda DNA) and that contain GC-rich sequences. 

Features

• 5’→3’ DNA polymerase activity 
• 3’→5’ exonuclease (proofreading) activity
• Suitable for GC-rich templates 
• High reaction rate: 7 seconds/kb 
• High fidelity: 70 times higher than Taq polymerase 
• Generates blunt end amplicons  
• Vast elongation capability (up to 40 kb) 
• Thermo-stable for more than 10 hrs at 95°C 

Storage

[TF3000] G-HiFi™ DNA Polymerase

-20°C for 24 months

The G-HiFi™ DNA Polymerase is a new genetically modified, recombinant DNA polymerase suitable for GC-rich templates that are difficult to amplify. The fidelity of G-HiFi™ DNA Polymerase is 70 times higher than that of Taq DNA polymerase. The high extension rate of G-HiFi™ DNA Polymerase is achieved by blending the DNA polymerase with an elongation enhancer. The optimized 5X G-HiFi™ Buffer includes special ingredients that suppress non-specific amplification as well as plateau effect produced by conventional PCR. With the optimized 5X G-HiFi™ Buffer, G-HiFi™ DNA Polymerase is capable to amplify most templates, such as longer targets (up to 40 kb from lambda DNA) and that contain GC-rich sequences.

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

RAA uses a novel RNA substrate tagged with a fluorescent reporter molecule (fluor) on one end and a quencher on the other. In the absence of RNases, the physical proximity of the quencher dampens fluorescence from the fluor to extremely low levels. When RNases are present, however, the RNA substrate is cleaved, and the fluor and quencher are spatially separated in solution. This causes the fluor to emit a bright green signal when excited by light of the appropriate wavelength. Fluorescence can be readily detected with a fluorometer. Since the fluorescence of the RAA Substrate increases over time when RNase activity is present, results monitored with a fluorometer can be evaluated kinetically. The sequence of the RAA Substrate has been carefully optimized to detect several RNases, including RNase A, RNase T1, RNase I, micrococcal nuclease, S1 nuclease, mung bean nuclease, and Benzonase.

RNase activity in a convenient and sensitive fluorimetric assay that delivers results in real time. Great for Quality Testing for RNase contamination of materials and supplies.