PrimePCR™ Probe Assay: NEB, 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: NEB, 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: NEB, 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:   qHsaCIP0028175
Assay Design:   Intron-spanning
Chromosome Location:   2:152362034-152362740question
Amplicon Length:   82
Splice Variants Targeted:   ENST00000409198 ENST00000397337 ENST00000413693 ENST00000434685 ENST00000421461 ENST00000424585 ENST00000397345 ENST00000427231 ENST00000172853 ENST00000397342 ENST00000509223

Gene Information

This gene encodes nebulin a giant protein component of the cytoskeletal matrix that coexists with the thick and thin filaments within the sarcomeres of skeletal muscle. In most vertebrates nebulin accounts for 3 to 4% of the total myofibrillar protein. The encoded protein contains approximately 30-amino acid long modules that can be classified into 7 types and other repeated modules. Protein isoform sizes vary from 600 to 800 kD due to alternative splicing that is tissue- species-and developmental stage-specific. Of the 183 exons in the nebulin gene at least 43 are alternatively spliced although exons 143 and 144 are not found in the same transcript. Of the several thousand transcript variants predicted for nebulin the RefSeq Project has decided to create three representative RefSeq records. Mutations in this gene are associated with recessive nemaline myopathy. [provided by RefSeq Sep 2009]

Gene Symbol:   NEB
Gene Name:   nebulin
Aliases:   DKFZp686C1456, FLJ11505, FLJ36536, FLJ39568, FLJ39584, NEB177D, NEM2
RefSeq:   NC_000002.11 NG_009382.2 NT_005403.17
Ensembl:   ENSG00000183091
Entrez:   4703
Chromosome Mapping:   2q22

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 0.999400
y-intercept 34.650000
Efficiency 98

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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