GeneAb™ MUC1

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Cod Uracil-DNA Glycosylase (Cod UNG) from Atlantic Cod is the only commercially available UNG enzyme that is completely and irreversibly inactivated by moderate heat treatment. The enzyme is produced in a recombinant E. coli (ung-) strain that contains a modified Cod UNG gene.

Key Features:

• Heat-labile – Completely and irreversibly inactivated at 55°C
• Contamination control – ideal in applications below
• Use of Cod UNG makes contamination control possible in RT-PCR
• Does not degrade PCR product post-PCR. This makes downstream use of the PCR product possible
• High purity enzyme, tested free of contaminating nucleases
• Detergent free
• Post-PCR Analysis – Enables post-PCR analysis

Applications

Ideal for contamination control in 

• PCR carry-over prevention
• RT-PCR
• RT-qPCR
• qPCR
• kPCR 
• RT-LAMP

Heat-labile Uracil-DNA Glycosylase

There are several commercially available Uracil-DNA glycosylases on the market today. Most of them are of bacterial origin and work well if you have no intention to further analyze the PCR products post-PCR. However, if you want to store your PCR products for downstream analysis such as cloning and sequencing, the reactivation of UNG and subsequent degradation of your PCR products are a problem with most of the commercially available UNGs. Cod UNG from ArcticZymes is completely and irreversibly inactivated by heat thus ensuring that sample integrity is maintained long-term regardless of storage conditions.

This is illustrated in figure 1, below

Figures

Properties

Recommended Protocols

1. Contamination control in PCR, qPCR and one-step RT-qPCR

Cod UNG works in all commercially available master mixes. 

Be sure that you have used dUTP containing dNTP mixes in your previous PCR experiments.

• Add 0.2 U Cod UNG directly to your 20 µl PCR reaction.
• pre-incubate for 5 min at room temperature.
• For RT-qPCR, reverse transcribe your RNA at 50-55°C.
• Run your PCR.
• Store your PCR product at -20°C or 4°C degrees.

2. Contamination control in RT-LAMP

Cod UNG is ideal for contamination control in RT-LAMP.  
One unit of Cod UNG per 30 μl reaction is sufficient for removing even high concentrations of carry-over contamination.

• Ensure that you use dNTP mixes containing dUTP in your experiments.
• Check that the RT-LAMP reaction is compatible with dUTP by running side-by-side reactions containing different ratios of dUTP to dUTP (100% dUTP, 90% dUTP, 80% dUTP and 0% dUTP).
• Add 1 U Cod UNG directly to your 30 µl RT-LAMP reaction.
• Prepare the reaction mix on ice.
• Analyze your RNA at 65°C, no preincubation is necessary.

Please refer to Protocol for Carry-over Contamination Control

Ordering Info

Cod Uracil-DNA Glycosylase (Cod UNG) from Atlantic Cod is the only commercially available UNG enzyme that is completely and irreversibly inactivated by moderate heat treatment. The enzyme is produced in a recombinant E. coli (ung-) strain that contains a modified Cod UNG gene.

Overview

• Quantify RNA of a wide spectrum of concentrations, including the lower ng per µL and pg per µL range
• Compatible with any Real-Time PCR system
• RNA is accurately quantified using a standard curve constructed from the provided RNA standard

Norgen’s Low Abundance RNA Quantification Kit offers a PCR-based detection procedure to quantify RNA of a wide spectrum of concentrations, including the lower ng per µL and pg per µL range. The kit has two main enzymatic components – reverse transcription using Norgen’s microScript Reverse Transcription system and PCR Master Mix used in conjunction with  a specially formulized primer mixture, to amplify human RNA from different types of inputs (such as various liquid biopsies). The kit is compatible with any Real-Time PCR system with the addition of fluorescent nucleic acid stains such as SYBR Green. The unknown RNA is accurately quantified by using a standard curve constructed from the provided RNA Standard. 

Details

Supporting Data

Figure 1 / 4

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Storage Conditions
Upon receipt, store Norgen’s Low Abundance RNA Quantification Kit at -20°C or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20°C or lower.

Component	Cat. 58900 (48 reactions)
microScript microRNA Enzyme Mix	20 μL
2x microScript Reverse Transcriptase Reaction Mix	0.5 mL
2X Real-Time PCR Master Mix	1 mL
RNA Quantification Primer Set Mix	200 μL
Quantified RNA Standard 50 pg/μL	2 x 60 μL
Nuclease-Free Water	3 x 1.25 mL
Product Insert	1

endo-BCN-PEG4-PFP ester is a click chemistry reagent containing a PFP ester and a BCN group. PFP esters have similar applications as the NHS esters, but are more stable in aqueous solution. They can be used to label the primary amines (-NH2) of proteins, amine-modified oligonucleotides, and other amine-containing molecules. The BCN group cais reactive with azide-tagged molecules. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

endo-BCN-PEG4-PFP ester is a click chemistry reagent containing a PFP ester and a BCN group. PFP esters have similar applications as the NHS esters, but are more stable in aqueous solution. They can be used to label the primary amines (-NH2) of proteins, amine-modified oligonucleotides, and other amine-containing molecules. The BCN group cais reactive with azide-tagged molecules. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.