PrimePCR™ Probe Assay: PHKB, 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: PHKB, 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: PHKB, 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:   qHsaCIP0031163
Assay Design:   Intron-spanning
Chromosome Location:   16:47621586-47622886question
Amplicon Length:   130
Splice Variants Targeted:   ENST00000323584 ENST00000299167 ENST00000455779

Gene Information

Phosphorylase kinase is a polymer of 16 subunits four each of alpha beta gamma and delta. The alpha subunit includes the skeletal muscle and hepatic isoforms encoded by two different genes. The beta subunit is the same in both the muscle and hepatic isoforms encoded by this gene which is a member of the phosphorylase b kinase regulatory subunit family. The gamma subunit also includes the skeletal muscle and hepatic isoforms encoded by two different genes. The delta subunit is a calmodulin and can be encoded by three different genes. The gamma subunits contain the active site of the enzyme whereas the alpha and beta subunits have regulatory functions controlled by phosphorylation. The delta subunit mediates the dependence of the enzyme on calcium concentration. Mutations in this gene cause glycogen storage disease type 9B also known as phosphorylase kinase deficiency of liver and muscle. Alternatively spliced transcript variants encoding different isoforms have been identified in this gene. Two pseudogenes have been found on chromosomes 14 and 20 respectively.[provided by RefSeq Feb 2010]

Gene Symbol:   PHKB
Gene Name:   phosphorylase kinase, beta
Aliases:   DKFZp781E15103, FLJ41698
RefSeq:   NC_000016.9 NG_016598.1 NT_010498.15
Ensembl:   ENSG00000102893
Entrez:   5257
Chromosome Mapping:   16q12-q13

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 0.998600
y-intercept 34.700000
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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