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Transcription Profiling of Malaria-Naïve and Semi-Immune Colombian Volunteers in a Plasmodium vivax Sporozoite Challenge [RT-PCR]

ABSTRACT: Whole blood gene expression was monitored in 85 samples using a Fluidigm nanoscale RT-qPCR array targeting 96 genes that we refer to as “blood informative transcripts” (BIT). There was no evidence for transcriptional changes prior to the appearance of blood stage parasite at day 12 or 13, at which time there was a strong interferon response and, unexpectedly, down-regulation of transcripts related to inflammation and innate immunity. This samples are related with the samples with accession number GSE67184 Reverse Transcription followed by quantitative PCR (RT-qPCR) was performed using Fluidigm 96×96 nanofluidic arrays targeting a set of 96 transcripts that are broadly informative of the major axes of variation for peripheral blood gene expression from Preininger et al. (2013) at six time-points (Pre-challenge, day 5, day 7, day 9, Diagnosis (Dx) and month 4).

ORGANISM(S): Homo sapiens

SUBMITTER: Monica Rojas-Peña

PROVIDER: E-GEOD-67469 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Transcription Profiling of Malaria-Naïve and Semi-Immune Colombian Volunteers in a Plasmodium vivax Sporozoite Challenge [RT-PCR]

Project description:Whole blood gene expression was monitored in 85 samples using a Fluidigm nanoscale RT-qPCR array targeting 96 genes that we refer to as “blood informative transcripts” (BIT). There was no evidence for transcriptional changes prior to the appearance of blood stage parasite at day 12 or 13, at which time there was a strong interferon response and, unexpectedly, down-regulation of transcripts related to inflammation and innate immunity. This samples are related with the samples with accession number GSE67184

2015-07-14 | GSE67469 | GEO

Single-cell profiling of mouse hematopoietic stem cells and their progeny

Project description:This dataset contains the single-cell gene expression profiles of mouse haematopoietic stem cell and progenitor cell populations isolated by FACS and paired gene expression profiles of their progeny (paired daughter cell assay and paired grand daughter cell assay). Expression of 96 TaqMan probes (A-MTAB-650) was assayed in individual cells by qRT-PCR using 96·96 Dynamic Arrays and the Biomark HD platform (Fluidigm).

2020-11-27 | E-MTAB-7504 | biostudies-arrayexpress

Real-time quantitative PCR analysis of 88 microRNAs involved in human cell differentiation and development in systemic sclerosis (SSc) dermal fibroblasts

Project description:Normal fibroblasts and SSc fibroblasts between the third and six subpassages were used for experiments. Total RNA was extracted from culture cells with ISOGEN (Nippon Gene, Tokyo, Japan). MicroRNA isolation from total RNA was performed using RT2 qPCR-Grade miRNA Isolation Kit (SA Bioscience). For RT2 Profiler PCR Array (SABioscience), microRNAs were reverse-transcribed into first strand cDNA using RT2 miRNA First Strand Kit (SABiosciences). A mixture of equal amounts of cDNAs from 5 normal fibroblasts or 5 SSc fibroblasts was prepared. The cDNA was mixed with RT2 SYBR Green/ROX qPCR Master Mix and the mixture was added into a 96-well RT2 miRNA PCR Array (SABiosciences) that included primer pairs for 88 human microRNAs. Human dermal fibroblasts were obtained by skin biopsy from the affected areas (dorsal forearm) of 5 patients with diffuse cutaneous SSc and <2 years of skin thickening. Control fibroblasts were obtained by skin biopsies from 5 healthy donors. Control donors were each matched with a SSc patient for age, sex, and biopsy site.

2012-01-01 | E-GEOD-34142 | biostudies-arrayexpress

Real-time quantitative PCR analysis of 88 microRNAs involved in human cell differentiation and development in normal fibroblasts stimulated with exogenous TGF-β1 and systemic sclerosis (SSc) dermal fibroblasts

Project description:Normal fibroblasts and SSc fibroblasts between the third and six subpassages were used for experiments. Normal and scleroderma fibroblasts were serum-starved for 24 hours and incubated in the presence or absence of TGF-β1 (2ng/ml) for 6 hours. Total RNA was extracted from culture cells with ISOGEN (Nippon Gene, Tokyo, Japan). MicroRNA isolation from total RNA was performed using RT2 qPCR-Grade miRNA Isolation Kit (SA Bioscience). For RT2 Profiler PCR Array (SABioscience), microRNAs were reverse-transcribed into first strand cDNA using RT2 miRNA First Strand Kit (SABiosciences). A mixture of equal amounts of cDNAs from 5 normal fibroblasts or 5 SSc fibroblasts was prepared. The cDNA was mixed with RT2 SYBR Green/ROX qPCR Master Mix and the mixture was added into a 96-well RT2 miRNA PCR Array (SABiosciences) that included primer pairs for 88 human microRNAs. Human dermal fibroblasts were obtained by skin biopsy from the affected areas (dorsal forearm) of 5 patients with diffuse cutaneous SSc and <2 years of skin thickening. Control fibroblasts were obtained by skin biopsies from 5 healthy donors. Control donors were each matched with a SSc patient for age, sex, and biopsy site.

2012-04-01 | E-GEOD-34827 | biostudies-arrayexpress

Heterogeneity in early lymphoid compartments [qPCR]

Project description:In order to understand the developmental trajectories of early lymphocyte development it is crucial to prospectively isolate stage and lineage specific cells. It has become clear that early lymphoid progenitor compartments in the bone marrow are molecularly and functionally heterogeneous which warrants further investigation to refine the marker combinations used to isolate these progenitors. An initial antibody screen revealed extensive surface marker heterogeneity amongst early lymphoid progenitors. This heterogeneity was resolved using single cell gene expression assays and single cell in vitro differentiation assays, identifying marker combinations that isolate functionally distinct populations. In addition, using reporter transgenic mice we were able to identify a set of surface markers that can be used alone or in combination with classical targets to identify specific stages of B-cell development. B-cell stages based on transgene expression which were used for screening purposes were verified by RNASeq. The data provides a greater resolution of the complexity of the lymphoid progenitor compartment within the bone marrow than has been understood to date and provides novel tools for the further identification of cell populations in B-lineage development. qPCR gene expression profiling of mouse common lymphoid progenitors (CLPs). Four 96-well plates with pre-amplified single CLP bone marrow cell cDNA (with 10 and 20 cell controls) were loaded on to a 96.96 Dynamic Array Chip for Gene Expression. Samples Ids follow where SS<X> denotes well ID, and P<X>denoted the plate. Controls include well where no RT is performed (noRT) and 10 and 20 cell controls.

2016-10-23 | E-GEOD-74606 | biostudies-arrayexpress

Homo sapiens

Project description:Blood-Informative Transcripts Define Nine Common Axes of Peripheral Blood Gene Expression

| PRJNA151861 | ENA

Fluidigm Biomark 96.96 nanofluidic arrays for qRT-PCR of 93 miRNAs: salivary extracellular vesicles

Project description:Crude extracellular vesicles (EVs) from eight healthy volunteers were separated into 6 fractions based on their densities by using the iodixanol-based density gradient centrifugation method. To determine the distribution of miRNAs among these fractions, quantities of 93 miRNAs were quantified by the TaqMan real time PCR method using the BioMark HD system (Fluidigm) equipped with 96.96 dynamic array (Fluidigm). Six samples were fractionated from a crude EVs by density gradient centrifugation. Total of 48 samples were prepared from 8 healthy volunteers. Technical replicate of 4 gave 8 x 6 x 4 x 93 = 17,856 data. As control Tris-HCl EDTA buffer (TE) was used.

2016-08-01 | E-GEOD-78763 | biostudies-arrayexpress

Real-time quantitative PCR analysis of 88 microRNAs involved in human cell differentiation and development in involved skin of systemic scleroderma (SSc), localized scleroderma (LSc) and Keloid.

Project description:Skin specimens were derived from involved skin of 3 systemic scleroderma (SSc), 3 localized scleroderma (LSc) and 3 keloid patients. These skin samples and 3 control skins were collected and fixed in formaldehyde immediately after resections. The microRNA (miRNA) isolation from human skin tissue was performed using miRNeasy FFPE kit (Qiagen). For PCR array, miRNAs were reverse-transcribed into first strand cDNA using RT2 miRNA First Strand Kit (SABiosciences). A mixture of equal amounts of miRNAs from 3 normal skins, 3 SSc, 3 LSc or 3 keloid were prepared, and miRNA expression profile in each disease in vivo was evaluated using RT2 Profiler PCR Array. The cDNA was mixed with RT2 SYBR Green/ROX qPCR Master Mix and the mixture was added into 96-well RT2 miRNA PCR Array that includes primer pairs for 88 human miRNAs (SABiosciences). Skin specimens were derived from involved skin of 3 systemic scleroderma (SSc), 3 localized scleroderma (LSc) and 3 keloid patients. These skin samples and 3 control skins were collected and fixed in formaldehyde immediately after resections. Control donors were each matched with diseases for age, sex, and biopsy site.

2013-01-15 | E-GEOD-43469 | biostudies-arrayexpress

A whole-blood RNA transcript based prognostic model in castration-resistant prostate cancer

Project description:Peripheral blood from 62 men with castration resistant prostate cancer was collected between 8/2006 and 6/2008. A panel of 168 inflammation-related and prostate cancer related genes was assessed with quantitative PCR to assess biomarkers predictive of survival. qPCR profiling of whole blood from patients with castration-resistant prostate cancer.

2013-04-02 | E-GEOD-45705 | biostudies-arrayexpress

Increased sensitivity of next generation sequencing-based expression profiling after globin reduction in human blood RNA

Project description:Transcriptome analysis is of great interest in clinical research, where significant differences between individuals can be translated into biomarkers of disease. Although next generation sequencing provides robust, comparable and highly informative expression profiling data, with several million of tags per blood sample, reticulocyte globin transcripts can constitute up to 76% of total mRNA compromising the detection of low abundant transcripts. We have removed globin transcripts from 6 human whole blood RNA samples with a human globin reduction kit and compared them with the same non-reduced samples using deep Serial Analysis of Gene Expression. Comparison of 6 whole blood RNA samples versus their globin reduced counterparts

2012-01-25 | E-GEOD-33701 | biostudies-arrayexpress

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