PrimePCR™ Probe Assay: CDKN2A, 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: CDKN2A, 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: CDKN2A, 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:   qHsaCEP0057827
Assay Design:   exonic
Chromosome Location:   9:21967834-21967949question
Amplicon Length:   86
Splice Variants Targeted:   ENST00000304494 ENST00000579755 ENST00000361570

Gene Information

This gene generates several transcript variants which differ in their first exons. At least three alternatively spliced variants encoding distinct proteins have been reported two of which encode structurally related isoforms known to function as inhibitors of CDK4 kinase. The remaining transcript includes an alternate first exon located 20 Kb upstream of the remainder of the gene; this transcript contains an alternate open reading frame (ARF) that specifies a protein which is structurally unrelated to the products of the other variants. This ARF product functions as a stabilizer of the tumor suppressor protein p53 as it can interact with and sequester MDM1 a protein responsible for the degradation of p53. In spite of the structural and functional differences the CDK inhibitor isoforms and the ARF product encoded by this gene through the regulatory roles of CDK4 and p53 in cell cycle G1 progression share a common functionality in cell cycle G1 control. This gene is frequently mutated or deleted in a wide variety of tumors and is known to be an important tumor suppressor gene. [provided by RefSeq Jul 2008]

Gene Symbol:   CDKN2A
Gene Name:   cyclin-dependent kinase inhibitor 2A (melanoma, p16, inhibits CDK4)
Aliases:   ARF, CDK4I, CDKN2, CMM2, INK4, INK4A, MLM, MTS-1, MTS1, P14, P14ARF, P16, P16-INK4A, P16INK4, P16INK4A, P19, P19ARF, TP16
RefSeq:   NC_000009.11 NG_007485.1 NT_008413.18
Ensembl:   ENSG00000147889
Entrez:   1029
Chromosome Mapping:   9p21

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 0.999400
y-intercept 36.930000
Efficiency 94

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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