PrimePCR™ Probe Assay: COL4A4, 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: COL4A4, 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: COL4A4, 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:   qHsaCIP0030571
Assay Design:   Intron-spanning
Chromosome Location:   2:227892631-227895207question
Amplicon Length:   114
Splice Variants Targeted:   ENST00000396625 ENST00000329662

Gene Information

This gene encodes one of the six subunits of type IV collagen the major structural component of basement membranes. This particular collagen IV subunit however is only found in a subset of basement membranes. Like the other members of the type IV collagen gene family this gene is organized in a head-to-head conformation with another type IV collagen gene so that each gene pair shares a common promoter. Mutations in this gene are associated with type II autosomal recessive Alport syndrome (hereditary glomerulonephropathy) and with familial benign hematuria (thin basement membrane disease). Two transcripts differing only in their transcription start sites have been identified for this gene and as is common for collagen genes multiple polyadenylation sites are found in the 3' UTR. [provided by RefSeq Jul 2008]

Gene Symbol:   COL4A4
Gene Name:   collagen, type IV, alpha 4
Aliases:   CA44
RefSeq:   NG_011591.1 NC_000002.11 NG_011592.1 NT_005403.17
Ensembl:   ENSG00000081052
Entrez:   1286
Chromosome Mapping:   2q35-q37

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 34.660000
Efficiency 101

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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