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
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.
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Cod UNG
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Product Info
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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
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.
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.
Mastitis is the single most costly disease of dairy cattle resulting in the reduction of milk yield and quality. The inflammation of the udder is mainly caused by infection of various bacteria. One of such mastitis bacteria, Streptococcus agalactiae, is highly infectious and causes mainly subclinical infections, which are not identified by the herdsman. As a result, S. agalactiae can spread widely within a herd, causing immediate loss due to reduced milk. S. agalactiae is a gram-positive bacteria belonging to the Group B streptococci. Traditional cultural identification of S. agalactiae is based on S. agalactiae being beta-hemolytic as well as presence of group B Lancefield antigen and by its ability to hydrolyze sodium hippurate.
Storage Conditions and Product Stability All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival.
