The genesig® Kit for Avian influenza A virus subtype (H5) (H5) genomes is designed for the in vitro quantification of H5 genomes. The kit is designed to have the broadest detection profile possible whilst remaining specific to the H5 genome. The primers and probe sequences in this kit have 100% homology with a broad range of H5 sequences based on a comprehensive bioinformatics analysis
The primers have very high homology with all reference sequences submitted since 2000 and therefore have the broadest detection profile. These primers will also detect most historical sequences for this subtype including H5N8, H5N2 and H5N9. However, due to the inherent instability of RNA viral genomes, novel emerging sequences may not be detected. Please contact us if you would like us to check the homology of our primers to specific strains you are working with
If you require further information, or have a specific question about the detection profile of this kit then please send an e.mail to [email protected] and our bioinformatics team will answer your question.
Rapid detection of all clinically relevant subtypes
Positive copy no. standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target
Accurate controls to confirm findings
ADVANCED KIT COMPONENTS:
H5 specific primer/probe mix (150 reactions; BROWN); FAM labeled
H5 positive control template (for Standard curve; RED)
Internal extraction control primer/probe mix (150 reactions; BROWN); VIC labeled as standard
Internal extraction control RNA (150 reactions; BLUE)
Endogenous control primer/probe mix (150 reactions; BROWN); FAM labeled
H5/Internal extraction control/endogenous control RT primer mix (150 reactions; GREEN). Required for two-step protocol only.
RNAse/DNAse free water (WHITE) for resuspension of primer/probe mixes and internal extraction control RNA
Template preparation buffer (YELLOW) for resuspension of internal control template, positive control template and standard curve preparation
KIT STORAGE AND STABILITY:This kit is stable at room temperature but should be stored at -20°C on arrival. Once the lyophilised components have been resuspended they should not be exposed to temperatures above -20°C for longer than 30 minutes at a time and unnecessary repeated freeze/thawing should be avoided. The kit is stable for six months from the date of resuspension under these circumstances
If a standard curve dilution series is prepared this can be stored frozen for an extended period. If you see any degradation in this serial dilution a fresh standard curve can be prepared from the positive control
We do not recommend using the kit after the expiry date stated on the pack.
REAGENTS AND EQUIPMENT TO BE SUPPLIED BY THE USER:
Real-Time PCR Instrument
RNA extraction kitThis kit is recommended for use with genesig Easy. DNA/RNA Extraction Kit. However, it is designed to work well with all processes that yield high quality DNA with minimal PCR inhibitors.
oasigTM lyophilized OneStep or Precision Plus OneStep 2X RT-qPCR Master Mix. Contains complete OneStep RT-qPCR master mix.
Pipettors and Tips
Vortex and centrifuge
Thin walled 1.5 ml PCR reaction tubes
SUITABLE SAMPLE MATERIAL:All kinds of sample material suited for PCR amplification can be used. Please ensure the samples are suitable in terms of purity, concentration, and RNA/DNA integrity (An internal PCR control is supplied to test for non specific PCR inhibitors). Always run at least one negative control with the samples. To prepare a negative-control, replace the template RNA sample with RNase/DNase free water.
DYNAMIC RANGE OF TEST:Under optimal PCR conditions genesig® etection kits have very high priming efficiencies of >95% and can detect less than 100 copies of target template.
INSTRUMENTATION REQUIREMENTS:Designed for use with real-time PCR instruments. . For use in genesig® q16, look for the “Easy” nomenclature.
ABOUT THE genesig® KITS:genesig® kits are sold for research use only and are not licensed for diagnostic procedures.
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, optimized and ready to use.
DNA/RNA EXTRACTION:genesig® Easy DNA/RNA extraction kits (sold separately) are a simple to use, low cost and powerful extraction technology suitable for virtually all sample types.
