PrimePCR™ Probe Assay: BEST3, 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: BEST3, 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: BEST3, 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:   qHsaCEP0051439
Assay Design:   exonic
Chromosome Location:   12:70072544-70072655question
Amplicon Length:   82
Splice Variants Targeted:   ENST00000331471 ENST00000488961 ENST00000330891 ENST00000553096 ENST00000476098 ENST00000552295 ENST00000548658

Gene Information

BEST3 belongs to the bestrophin family of anion channels which includes BEST1 (MIM 607854) the gene mutant in vitelliform macular dystrophy (VMD; MIM 153700) and 2 other BEST1-like genes BEST2 (MIM 607335) and BEST4 (MIM 607336). Bestrophins are transmembrane (TM) proteins that share a homology region containing a high content of aromatic residues including an invariant arg-phe-pro (RFP) motif. The bestrophin genes share a conserved gene structure with almost identical sizes of the 8 RFP-TM domain-encoding exons and highly conserved exon-intron boundaries. Each of the 4 bestrophin genes has a unique 3-prime end of variable length (Stohr et al. 2002 [PubMed 12032738]; Tsunenari et al. 2003 [PubMed 12907679]).[supplied by OMIM Mar 2008]

Gene Symbol:   BEST3
Gene Name:   bestrophin 3
Aliases:   MGC13168, MGC40411, VMD2L3
RefSeq:   NC_000012.11 NT_029419.12
Ensembl:   ENSG00000127325
Entrez:   144453
Chromosome Mapping:   12q14.2-q15

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 35.630000
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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