PrimePCR™ Probe Assay: SHH, 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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Assay Information

Technology:   qPCR
Assay Type:   Probe
Application:   Gene Expression
Unique Assay ID:   qHsaCEP0040459
Assay Design:   exonic
Chromosome Location:   7:155599083-155604529question
Amplicon Length:   151
Splice Variants Targeted:   ENST00000297261 ENST00000430104

Gene Information

This gene encodes a protein that is instrumental in patterning the early embryo. It has been implicated as the key inductive signal in patterning of the ventral neural tube the anterior-posterior limb axis and the ventral somites. Of three human proteins showing sequence and functional similarity to the sonic hedgehog protein of Drosophila this protein is the most similar. The protein is made as a precursor that is autocatalytically cleaved; the N-terminal portion is soluble and contains the signalling activity while the C-terminal portion is involved in precursor processing. More importantly the C-terminal product covalently attaches a cholesterol moiety to the N-terminal product restricting the N-terminal product to the cell surface and preventing it from freely diffusing throughout the developing embryo. Defects in this protein or in its signalling pathway are a cause of holoprosencephaly (HPE) a disorder in which the developing forebrain fails to correctly separate into right and left hemispheres. HPE is manifested by facial deformities. It is also thought that mutations in this gene or in its signalling pathway may be responsible for VACTERL syndrome which is characterized by vertebral defects anal atresia tracheoesophageal fistula with esophageal atresia radial and renal dysplasia cardiac anomalies and limb abnormalities. Additionally mutations in a long range enhancer located approximately 1 megabase upstream of this gene disrupt limb patterning and can result in preaxial polydactyly. [provided by RefSeq Jul 2008]

Gene Symbol:   SHH
Gene Name:   sonic hedgehog
Aliases:   HHG1, HLP3, HPE3, MCOPCB5, SMMCI, TPT, TPTPS
RefSeq:   NC_000007.13 NG_007504.1 NT_007741.14
Ensembl:   ENSG00000164690
Entrez:   6469
UniGene:   Hs.164537
Chromosome Mapping:   7q36

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 0.999800
y-intercept 36.760000
Efficiency 95

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.

Number Description Download
6262 PrimePCR Assays: Meeting the MIQE Guidelines by Full Wet-Lab Validation, Rev B Click to download
6263 PrimePCR Pathway Analysis: Pathway Curation and Real-Time PCR Panel Design Strategy, Rev B Click to download
6290 PrimePCR Assays and Panels Brochure, Rev D Click to download
LIT10026370 Instruction Manual, PrimePCR Assays, Panels, and Controls, Rev E Click to download
10039761 PrimePCR™ Assays Quick Guide, Ver B Click to download