PrimePCR™ Probe Assay: PHKA1, 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: PHKA1, 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: PHKA1, 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:   qHsaCEP0049515
Assay Design:   exonic
Chromosome Location:   X:71800752-71800898question
Amplicon Length:   117
Splice Variants Targeted:   ENST00000373542 ENST00000373545 ENST00000373539 ENST00000339490 ENST00000541944 ENST00000593378 ENST00000598643 ENST00000600891 ENST00000601371 ENST00000596491

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 and the skeletal muscle isoform is encoded by this gene. The beta subunit is the same in both the muscle and hepatic isoforms and encoded by one gene. The gamma subunit also includes the skeletal muscle and hepatic isoforms which are 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 9D also known as X-linked muscle glycogenosis. Alternatively spliced transcript variants encoding different isoforms have been identified in this gene. A pseudogene has been found on chromosome 1.[provided by RefSeq Feb 2010]

Gene Symbol:   PHKA1
Gene Name:   phosphorylase kinase, alpha 1 (muscle)
Aliases:   MGC132604, PHKA
RefSeq:   NC_000023.10 NT_011669.17 NG_016599.1
Ensembl:   ENSG00000067177
Entrez:   5255
Chromosome Mapping:   Xq12-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™ Universal SYBR® Green Supermix
Experimental Sample qPCR Human Reference Total RNA

Summary Data:

R2 0.999300
y-intercept 35.210000
Efficiency 102

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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