PrimePCR™ Probe Assay: ROBO3, Human

RT

Real-time PCR probe assay designed for gene expression analysis. Probe assays consist of unlabeled PCR primers and a dual labeled fluorescent probe.

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PrimePCR™ PreAmp for Probe Assay: ROBO3, Human
Reaction:     400 reactions
Gene-specific PCR primers for the unbiased preamplification of small quantities of cDNA for subsequent use in downstream gene expression analysis.

PrimePCR™ Template for Probe Assay: ROBO3, Human
Reaction:     200 x 20 µl reactions
Gene-specific synthetic DNA template designed to give a positive real-time PCR result when used with the corresponding probe assay.


Assay Information

Technology:   qPCR
Assay Type:   Probe
Application:   Gene Expression
Unique Assay ID:   qHsaCIP0033431
Assay Design:   Intron-spanning
Chromosome Location:   11:124746329-124747435question
Amplicon Length:   85
Splice Variants Targeted:   ENST00000397801 ENST00000538940

Gene Information

This gene is a member of the Roundabout (ROBO) gene family that controls neurite outgrowth growth cone guidance and axon fasciculation. ROBO proteins are a subfamily of the immunoglobulin transmembrane receptor superfamily. SLIT proteins 1-3 a family of secreted chemorepellants are ligands for ROBO proteins and SLIT/ROBO interactions regulate myogenesis leukocyte migration kidney morphogenesis angiogenesis and vasculogenesis in addition to neurogenesis. This gene ROBO3 has a putative extracellular domain with five immunoglobulin (Ig)-like loops and three fibronectin (Fn) type III motifs a transmembrane segment and a cytoplasmic tail with three conserved signaling motifs: CC0 CC2 and CC3 (CC for conserved cytoplasmic). Unlike other ROBO family members ROBO3 lacks motif CC1. The ROBO3 gene regulates axonal navigation at the ventral midline of the neural tube. In mouse loss of Robo3 results in a complete failure of commissural axons to cross the midline throughout the spinal cord and the hindbrain. Mutations ROBO3 result in horizontal gaze palsy with progressive scoliosis (HGPPS); an autosomal recessive disorder characterized by congenital absence of horizontal gaze progressive scoliosis and failure of the corticospinal and somatosensory axon tracts to cross the midline in the medulla. Alternative transcript variants have been described but have not been experimentally validated. [provided by RefSeq Dec 2009]

Gene Symbol:   ROBO3
Gene Name:   roundabout, axon guidance receptor, homolog 3 (Drosophila)
Aliases:   FLJ21044, HGPPS, HGPS, RBIG1, RIG1
RefSeq:   NC_000011.9 NG_016214.1 NT_033899.8
Ensembl:   ENSG00000154134
Entrez:   64221
Chromosome Mapping:   11q24.2

The below validation information is for the Primer Pair only   Download Validation Data (.pdf)

Products used to generate validation data:

Real-Time PCR Instrument CFX384 Real-Time PCR Detection System
Reverse Transcription Reagent iScript™ Advanced cDNA Synthesis Kit for RT-qPCR
Real-Time PCR Supermix SsoAdvanced™ SYBR® Green Supermix
Experimental Sample qPCR Human Reference Total RNA

Summary Data:

R2 0.999000
y-intercept 35.190000
Efficiency 96

Amplification Plot
Amplification of cDNA generated from universal RNA.

Amplification of cDNA generated from universal RNA.

Melt Peak
Melt curve analysis of above amplification.

Melt curve analysis of above amplification.

Standard Curve
Standard curve generated using 20 million copies of template diluted 10 fold to 20 copies.

Standard curve generated using 20 million copies of template diluted 10 fold to 20 copies.

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