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The validated multiplex real-time PCR was used to detect and differentiate BoHV-1 and BoHV-5 in bovine clinical samples with known histories. TaqMan MGB, TaqMan probes, Molecular Beacons and real time PCR primers. You will now be able to design thousands of efficient, highly specific oligos to make microarrays for SNP genotyping and expression studies with the click of a button. The assay specificity was demonstrated by testing bacterial and viral DNA from pathogens commonly isolated from bovine respiratory and reproductive tracts. Microarray Probe Design for Gene Expression and SNP Genotyping- Version 2.00 of AlleleID® includes a separate module for supporting high throughput microarray probe design. Standard curves relating Ct value to template copy number had correlation coefficients of 0.989 for BoHV-1 and 0.978 for BoHV-5.
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#ALLELEID MGB ALLELE PROBE SOFTWARE#
The software parameters were set to avoid cross-homology with the equine genome and cross-hybridization interference with the adopted oligonucleotides specific to the 18S rRNA gene of B. The multiplex real-time PCR had reaction efficiencies of 1.04 for BoHV-1 and 1.08 for BoHV-5. Primers and a minor groove binder (MGB) hydrolysis probe were designed using the AlleleID 7.72 software (Premier Biosoft, Palo Alto, CA, USA). The detection limits of the duplex assay were 10 copies of BoHV-1 and 45 copies of BoHV-5. BoHV-1 and BoHV-5 were also correctly identified when incorporated in spiked semen and brain tissue samples. The assay correctly identified 22 BoHV-1 and six BoHV-5 isolates from the Biosecurity Sciences Laboratory virus collection. The multiplex real-time PCR combines a duplex real-time PCR that targets the DNA polymerase gene of BoHV-1 and BoHV-5 and a real-time PCR targeting mitochondrial DNA, as a house-keeping gene, described previously by Cawthraw et al. KRAS G12R assays comprised of MGB Eclipse Probes (FAM dyewild-type probe TET dyemutant probe) and primers manufactured by IDT or assays for Research Use Only were used. Equivalent results for assays with IDT MGB Eclipse Probes and assays for Research Use Only.
#ALLELEID MGB ALLELE PROBE HOW TO#
pronouncekiwi - How To Pronounce AlleleID. Thank you for helping build the largest language community on the internet. A multiplex real-time PCR was developed for the detection and differentiation of two closely related bovine herpesviruses 1 (BoHV-1) and 5 (BoHV-5). Allele 1 - FAM (RFU) Genotyping calls Figure 2. Listen to the audio pronunciation of AlleleID on pronouncekiwi.