Kinetic PCR analysis using a CCD camera and without using oligonucleotide probes

Higuchi, Russell; Watson, Robert H.

doi:10.1016/b978-012372185-3/50017-1

Cited by 16 publications

(11 citation statements)

References 17 publications

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“…We also analyzed mRNA expression of a selected set of genes in Th1 and Th2 RNA samples using kinetic RT-PCR. 47 As expected, we noticed variability in gene expression changes in cell lines derived from different subjects, but we could confirm differential expression of 28 out of 29 genes in Th1 and Th2 cells generated from two independent donors. Well-established marker genes for Th1 cells, such as IFN-γ and IL-12Rβ2 were found at much higher levels in Th1 than in Th2 cells.…”

Section: Transcript Imagining Of Human and Mouse T Helper Cell Subsetssupporting

confidence: 74%

A Genomic View of Helper T Cell Subsets

Rogge

2002

Annals of the New York Academy of Sciences

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Genomic-scale gene expression profiling in combination with the availability of a draft sequence of the human genome is beginning to revolutionize the way immunology is done. The possibility of measuring levels of gene expression for tens of thousands of genes simultaneously and in a quantitative fashion aids in the definition of a comprehensive molecular phenotype of cells and cellular processes of the immune system in health and disease. T helper lymphocytes are an essential element of appropriate immune responses to pathogens. To achieve effective immunity, T helper cells differentiate into at least two specialized subsets that direct type 1 and type 2 immune responses. Here, I discuss recent progress that has been made in our understanding of the genetic program that controls the development and functional properties of helper T cell subsets.

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Section: Transcript Imagining Of Human and Mouse T Helper Cell Subsetssupporting

confidence: 74%

A Genomic View of Helper T Cell Subsets

Rogge

2002

Annals of the New York Academy of Sciences

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“…A one-cycle delay (ΔCt=1) indicates that the ratio of one allele to the other is approximately 1:2; a two-cycle delay (ΔCt=2), 1:4; or in general, 1 : 2 ÁCt if amplification efficiency is very close to 100%. Under optimized PCR conditions, PCR amplification efficiency is very close to 100% (Higuchi and Watson 1999), and the exponential parameter is 2. However, when PCR conditions are adjusted (mainly by increasing annealing/extension temperature) to better discriminate allele-specific signals, the efficiency of amplification may be slightly less than 2.…”

Section: Principle and Methods Of Computation Of Allele Expression Ratiosmentioning

confidence: 93%

Differential expression of human COMT alleles in brain and lymphoblasts detected by RT-coupled 5? nuclease assay

et al. 2004

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“…Our original method required hybridization of PCR‐amplified product with radioactive probes, autoradiography, and image analysis, with related safety concerns and expense 14,15 . We report here the application of another PCR detection method, the kinetic (or real‐time quantitative) PCR (kPCR), 16‐18 to achieve high throughput quantitation of as little as 0.008 WBCs per μL of filtered blood. Specifically, we have optimized the assay targeting a highly conserved region of HLA DQα gene, which is present in two copies per WBC but not present in RBCs or platelets.…”

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confidence: 99%