Paxgene blood mrna kit

Manufactured by Preanalytix

Sourced in Switzerland

The PAXgene Blood mRNA Kit is a lab equipment product that is used for the stabilization and purification of RNA from whole blood samples. The kit provides a standardized method for collecting, storing, and processing blood samples to obtain high-quality RNA for further analysis.

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Lab products found in correlation

Paxgene blood rna tube, by Preanalytix (2 mentions) Superscript 3 first strand synthesis system, by Thermo Fisher Scientific (2 mentions) 2100 bioanalyzer, by Agilent Technologies (2 mentions) Allprep dna rna mirna universal kit, by Qiagen (2 mentions) Wt ovation pico rna amplification system, by Tecan (1 mentions) Netaffx analysis center version 31, by Thermo Fisher Scientific (1 mentions) Human exon 1.0 st array, by Thermo Fisher Scientific (1 mentions) Paxgene rna tube, by Qiagen (1 mentions) Nanodrop nd 1000 spectrophotometer, by Thermo Fisher Scientific (1 mentions) Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions) Nanodrop 2000 spectrophotometry, by Thermo Fisher Scientific (1 mentions) Trizol ls, by Thermo Fisher Scientific (1 mentions) 2100 bioanalyzer rna 6000 nano labchip, by Agilent Technologies (1 mentions) Rnase free dnase 1, by Thermo Fisher Scientific (1 mentions)

8 protocols using paxgene blood mrna kit

RNA Extraction and Reverse Transcription

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RNA was extracted from blood collected in PAXgene Blood RNA tubes (PreAnalytix GmbH, Qiagen, Germantown, MD) and stored at −80°C. Total RNA was extracted using the PAXgene Blood mRNA Kit (PreAnalytix GmbH, Qiagen Sciences Inc, Germantown, MD) following the manufacturer’s protocol.
The RNA from the NAc core was extracted using the All Prep DNA/RNA/miRNA Universal kit (Qiagen Sciences, Inc., Germantown, MD) following the manufacturer’s instructions.
RNA quantity and quality was evaluated on a 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA). SuperScript III First-strand Synthesis System (Life Technologies, Carlsbad, CA) was used to reverse-transcribe 500 ng of each RNA sample following the manufacturer’s instructions.

Cervera-Juanes R., Wilhem L.J., Park B., Lee R., Locke J., Helms C., Gonzales S., Wand G., Jones S.R., Grant K.A, & Ferguson B. (2015). MAOA EXPRESSION PREDICTS VULNERABILITY FOR ALCOHOL USE. Molecular psychiatry, 21(4), 472-479.

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NF-κB Gene Expression Analysis

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Gene expression analysis of a predefined group of 200 NF-κB-regulated genes (Broad Institute curated HALLMARK NF-κB gene-set [39 (link)]) was evaluated via mRNA Sequencing (Illumina) on RNA isolated from whole blood (PAXgene Blood mRNA Kit, PreAnalytix).

Finanger E., Vandenborne K., Finkel R.S., Lee Sweeney H., Tennekoon G., Yum S., Mancini M., Bista P., Nichols A., Liu H., Fretzen A, & Donovan J.M. (2019). Phase 1 Study of Edasalonexent (CAT-1004), an Oral NF-κB Inhibitor, in Pediatric Patients with Duchenne Muscular Dystrophy. Journal of Neuromuscular Diseases, 6(1), 43-54.

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Profiling Gene Expression in Blood

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The methods for gene expression profiling were previously published (Joehanes and others 2013 ). Briefly, peripheral blood samples were extracted using the PAXgene Blood mRNA kit (PreAnalytiX, Hombrechtikon, Switzerland), and amplified by the WT-Ovation Pico RNA Amplification System (NuGEN, San Carlos, CA), according to manufacturer’s instructions. cDNA was then hybridized to the Human Exon 1.0 ST Array (Affymetrix, Inc., Santa Clara, CA) for quantification. The raw data were quantile-normalized and natural-log transformed, followed by summarization using Robust Multi-array Average (Irizarry and others 2003 (link)). The gene annotations were obtained from Affymetrix NetAffx Analysis Center (version 31). We excluded transcript clusters that were not mapped to RefSeq transcripts, resulting in 17,873 distinct transcripts (17,324 unique gene identifiers) for downstream analysis.

Pilling L.C., Joehanes R., Melzer D., Harries L.W., Henley W., Dupuis J., Lin H., Mitchell M., Hernandez D., Ying S.X., Lunetta K.L., Benjamin E.J., Singleton A., Levy D., Munson P., Murabito J.M, & Ferrucci L. (2015). Gene Expression markers of Age-Related Inflammation in Two Human Cohorts. Experimental gerontology, 70, 37-45.

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RNA Extraction from Blood Samples

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Blood samples were collected at the visit during which the routine glucose challenge test was taken, approximately between 24 and 28 weeks gestation. Blood was drawn into a 2.5 mL Qiagen PAXgene RNA tube, containing an RNA stabilizing agent. Collection tubes were maintained at 4 °C and sent to the study laboratory at Dartmouth within 24 h and stored at −80 °C until they were processed for RNA isolation. Samples were thawed and processed in batches using the PAXgene Blood mRNA kit (PreAnalytix 763134), according to the manufacturer’s instructions, and quantified by Nanodrop. In order to remove any contaminating DNA, RNase-free DNase was added during the extraction process. RNA integrity of batch controls was assessed by BioAnalyzer fragment analysis.

Bommarito P.A., Martin E., Smeester L., Palys T., Baker E.R., Karagas M.R, & Fry R.C. (2017). Fetal-sex dependent genomic responses in the circulating lymphocytes of arsenic-exposed pregnant women in New Hampshire. Reproductive toxicology (Elmsford, N.Y.), 73, 184-195.

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Extracting High-Quality RNA from Venipuncture Blood

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Total RNA from venipuncture PAXgene blood collection tubes was extracted using the PAXgene Blood mRNA kit (PreAnalytiX GmbH, Switzerland) according to the manufacturer’s protocol. The RNA yield was determined by a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA), while the quality and integrity of the RNA were assessed on an Agilent 2100 BioAnalyzer (Agilent Technologies, Amstelveen, The Netherlands) using the RNA 6000 Nano Chip kit. The average RNA integrity number of the total RNA samples obtained from PAXgene tubes was 9.6 ± 0.05.

Weinberger B., Haks M.C., de Paus R.A., Ottenhoff T.H., Bauer T, & Grubeck-Loebenstein B. (2018). Impaired Immune Response to Primary but Not to Booster Vaccination Against Hepatitis B in Older Adults. Frontiers in Immunology, 9, 1035.

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RNA Extraction and Reverse Transcription

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Evaluating NF-κB Pathway Modulation

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NF-κB inhibition was evaluated by measuring the expression of genes whose transcription is directly regulated by NF-κB activation [32] .
Target genes included the alternative NF-κB pathway component p100 (also known as NFKB2 ) that contains 6 NF-κB binding sites in the promoter region, and the inhibitor of NF-κB, NFKBIA (aka IkappaBalpha) that contains 3 NF-κB binding sites in promoter region. Gene expression was evaluated via mRNA Sequencing (Illumina) on RNA isolated from whole blood (PAXgene Blood mRNA Kit, PreAnalytix) collected at baseline, and weeks 4, 12 and 24.
Blood samples for evaluation of biomarkers of inflammation and muscle damage, including a cytokine panel of multiple inflammatory markers were collected predose at baseline, then every 4 weeks through week 16, then every 12 weeks through week 72.
Heart rate was measured following a rest period as part of ECG collection.

Finkel R.S., Finanger E., Vandenborne K., Sweeney H.L., Tennekoon G., Shieh P.B., Willcocks R., Walter G., Rooney W.D., Forbes S.C., Triplett W.T., Yum S.W., Mancini M., MacDougall J., Fretzen A., Bista P., Nichols A, & Donovan J.M. (2021). Disease-modifying effects of edasalonexent, an NF-κB inhibitor, in young boys with Duchenne muscular dystrophy: Results of the MoveDMD phase 2 and open label extension trial. Neuromuscular disorders : NMD, 31(5).

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Aortic Valve RNA Extraction Workflow

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For each valve, total RNA from a single aortic valve cusp was extracted using the TRIzol LS (Invitrogen, Carlsbad, CA, USA) method and digested with RNase free DNase I (Invitrogen).
For PESA and CAVD validation total RNA was using either the PAXgene blood mRNA kit (PreAnalytiX, Hombrechtikon, Switzerland) for manual isolation or the QIAsymphony PAXgene blood RNA kit (PreAnalytiX) for automated isolation using a QIAsymphony SP liquid handling robot (Qiagen, Venlo, Netherlands). RNA purity, concentration, and integrity were assessed by Nanodrop 2000 spectrophotometry (Thermo Scientific) and automated electrophoresis in a 2100
Bioanalyzer (RNA6000 Nano LabChip; Agilent). Samples with a RIN > 6 were selected for reverse transcription.

MacGrogan D., Martínez-Poveda B., Desvignes J.P., Fernandez-Friera L., Gomez M.J., Gil Vilariño E., Callejas Alejano S., Garcia-Pavia P., Solis J., Lucena J., Salgado D., Collod-Béroud G., Faure E., Théron A., Torrents J., Avierinos J.F., Montes L., Dopazo A., Fuster V., Ibañez B., Sánchez-Cabo F., Zaffran S, & de la Pompa J.L. (2020). Identification of a peripheral blood gene signature predicting aortic valve calcification. Physiological genomics, 52(12).

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