PrimePCR™ Probe Assay: HLA-A, 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: HLA-A, 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: HLA-A, 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:   qHsaCEP0040111
Assay Design:   exonic
Chromosome Location:   6:29913295-29913439question
Amplicon Length:   115
Splice Variants Targeted:   ENST00000564856 ENST00000396634 ENST00000376806 ENST00000376802 ENST00000376809 ENST00000457879 ENST00000438861 ENST00000453975 ENST00000547271 ENST00000453057 ENST00000455882 ENST00000413609 ENST00000547522 ENST00000431930 ENST00000444289 ENST00000416096 ENST00000552193 ENST00000551120 ENST00000443552 ENST00000417978 ENST00000442939 ENST00000549869 ENST00000552498 ENST00000438447 ENST00000282493 ENST00000383605 ENST00000383619 ENST00000376822 ENST00000549224 ENST00000456012 ENST00000450342 ENST

Gene Information

HLA-A belongs to the HLA class I heavy chain paralogues. This class I molecule is a heterodimer consisting of a heavy chain and a light chain (beta-2 microglobulin). The heavy chain is anchored in the membrane. Class I molecules play a central role in the immune system by presenting peptides derived from the endoplasmic reticulum lumen. They are expressed in nearly all cells. The heavy chain is approximately 45 kDa and its gene contains 8 exons. Exon 1 encodes the leader peptide exons 2 and 3 encode the alpha1 and alpha2 domains which both bind the peptide exon 4 encodes the alpha3 domain exon 5 encodes the transmembrane region and exons 6 and 7 encode the cytoplasmic tail. Polymorphisms within exon 2 and exon 3 are responsible for the peptide binding specificity of each class one molecule. Typing for these polymorphisms is routinely done for bone marrow and kidney transplantation. Hundreds of HLA-A alleles have been described. [provided by RefSeq Jul 2008]

Gene Symbol:   HLA-A
Gene Name:   major histocompatibility complex, class I, A
Aliases:   FLJ26655, HLAA
RefSeq:   NC_000006.11 NT_007592.15 NT_113891.2 NT_167244.1
Ensembl:   ENSG00000206503
Entrez:   3105
Chromosome Mapping:   6p21.3

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 0.999600
y-intercept 35.080000
Efficiency 100

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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