CO00016 Midstream Cassette 140x15mm, blue w/lens

Overview

• Detection kits for HPV (High and Low Risk)
• Lyophilized format designed for room temperature shipping
• Available in TaqMan format for analysis

Norgen’s HPV (High and Low Risk) TaqMan PCR Lyophilized Kit is designed for the detection of HPV High and Low Risk specific DNA in a real-time PCR based on the use of TaqMan® technology. The lyophilized format is designed to ship the kit at ambient temperature.

Norgen’s HPV (High and Low Risk) TaqMan Lyophilized Probe/Primer and Control Set is designed for the detection of HPV High and Low Risk specific DNA in a real-time PCR based on the use of TaqMan® technology. The lyophilized format is designed to ship the kit at ambient temperature.

Details

Supporting Data

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Storage Conditions and Product Stability

• All kit components should be stored at -20°C upon arrival.
• Once reconstituted, repeated thawing and freezing (>2 times) 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 kit components can be stored for 2 years after the date of production without showing any reduction in performance.

OverView

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

• Isolate all sizes of circulating and exosomal RNA, including microRNA
• Isolate all sizes of circulating DNA from plasma and serum samples
• Isolate viral and bacterial DNA and RNA
• Versatile plasma and serum input volumes (10 µL – 200 µL)
• No phenol extractions
• Bind and elute all RNA irrespective of size or GC content, without bias
• Concentrate circulating RNA, exosomal RNA and cell-free circulating DNA into a flexible elution volume ranging from 10 µL to 25 µL
• Isolate inhibitor-free nucleic acids
• Purify high-quality RNA and DNA in 30 minutes
• Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

Norgen’s Plasma/Serum RNA/DNA Purification Mini Kit provides a fast, reliable, reproducible and simple procedure for the sequential isolation of circulating RNA, exosomal RNA and Cell-Free Circulating DNA (cfc) from the same Plasma/Serum sample. It can sequentially isolated RNA and DNA from small plasma/serum input ranging from 10 µL to 200 µL. The kit is designed to isolate all sizes of circulating RNA, including microRNA, all sizes of exosomal RNA as well as all sizes of cfc-DNA from fresh or frozen plasma or serum samples. Norgen’s Plasma/Serum RNA Purification Kit provides a clear advantage over other available kits in that it does not require Phenol/Chloroform or any Protease treatments for the isolation of plasma/serum RNA or DNA . RNA and DNA can be eluted into a flexible elution volume ranging from 10 µL to 25 µL. Purified RNA can be used in a number of sensitive downstream applications including reverse transcription qPCR, reverse transcription PCR, NGS, Northern blotting, RNase protection and primer extension, and expression array assays. Purified DNA can be used in a number of sensitive downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis.

Background

Typical yields of free-circulating, exosomal RNA and cfc-DNA vary depending on the input sample, as the amount of RNA and/or DNA present in plasma and serum will depend upon the health status of the individual.  Normally, the RNA/DNA yield from plasma or serum is highly variable (range from 1 to 100 ng/mL). Variability is also observed between samples collected from the same donor at different times during the day. This kit is suitable for the isolation of RNA/DNA from serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.

Details

Supporting Data

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 Note: Do not exceed the recommended sample input volume of 200 µL.

Storage Conditions and Product Stability

All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.