Paxgene blood mirna kit

Manufactured by Preanalytix

Sourced in Switzerland, Germany

The PAXgene Blood miRNA kit is a laboratory product designed to facilitate the collection, stabilization, and purification of microRNA (miRNA) from whole blood samples. The kit provides a standardized procedure for the extraction and isolation of miRNA, which can be used for subsequent analysis and research purposes.

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Lab products found in correlation

Paxgene blood rna tube, by Preanalytix (16 mentions) Agilent 2100 bioanalyzer, by Agilent Technologies (7 mentions) Qiacube, by Qiagen (4 mentions) Mirneasy mini kit, by Qiagen (4 mentions) Qubit fluorometer, by Thermo Fisher Scientific (3 mentions) Rna 6000 nano kit, by Agilent Technologies (2 mentions) 2100 bioanalyzer, by Agilent Technologies (2 mentions) Agilent 2200 tapestation system, by Agilent Technologies (2 mentions) Nanodrop 1000, by Thermo Fisher Scientific (2 mentions) Nanodrop nd 1000 spectrophotometer, by Thermo Fisher Scientific (2 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (2 mentions) Viia7 real time pcr thermal cycler, by Thermo Fisher Scientific (2 mentions) Magmax for stabilized blood tubes rna isolation kit, by Thermo Fisher Scientific (2 mentions) Genetitan mc scanner, by Thermo Fisher Scientific (2 mentions) Rna broad range assay kit, by Thermo Fisher Scientific (2 mentions) Allprep dna rna mirna universal kit, by Qiagen (1 mentions) Agilent bioanalyzer, by Agilent Technologies (1 mentions) Globinclear, by Thermo Fisher Scientific (1 mentions) Minelute rna cleanup kit, by Qiagen (1 mentions) Nd 1000 spectrophotometer, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Blood miRNA Kit (3 citations)

40 protocols using paxgene blood mirna kit

Transcriptome Profiling of Whole Blood and Separated Cells

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In addition to the subset of individuals with detailed separate cell samples (n=60), a further eight patients were included in gene expression array analyses (total n=68). Separated cell RNA was extracted using the Allprep DNA/RNA miRNA universal kit (Qiagen). Whole blood RNA was extracted from PAXgene tubes using the PAXgene blood miRNA kit (PreAnalytix, Switzerland). The RNA was quantified and assessed for quality using the Agilent BioAnalyzer with only samples with a RNA-integrity number of >7 being used for downstream analyses. Following sample concentration and cleanup using the MinElute RNA cleanup kit (Qiagen), globin mRNA transcripts were depleted using GlobinClear (Ambion, Life Technologies USA). RNA was amplified and biotylated using the Illumina TotalPrep RNA Amplification Kit (Ambion, Life Technology). The cRNA was quantified and assessed for quality using the Agilent BioAnalyzer with the expected gel appearance of cRNA is a ‘smear', with a distribution of cRNA size is expected between 250–5,500 nucleotides, with most cRNA between 1,000 and 1,500 nt. Illumina HT12 human v4 expression microarrays were performed using a hybridization time of 18 h at 58 °C. Data were analysed using the lumi55 (link) and limma61 packages. Data were background adjusted, variance stabilized and quantile normalized.

Ventham N.T., Kennedy N.A., Adams A.T., Kalla R., Heath S., O'Leary K.R., Drummond H., Wilson D.C., Gut I.G., Nimmo E.R, & Satsangi J. (2016). Integrative epigenome-wide analysis demonstrates that DNA methylation may mediate genetic risk in inflammatory bowel disease. Nature Communications, 7, 13507.

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Whole Blood RNA Profiling for Transcriptomics

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Total RNA was extracted from PAXgene blood on the QIAcube (Qiagen, Germany), using the PAXgene Blood miRNA kit from PreAnalytix, according to manufacturer’s instructions. Human Gene 1.1 (GPL6244; RTP and CHFP cohorts) ST array plates (Affymetrix, United States) were used to measure mRNA abundance. This work was carried out at The Scripps Research Institute DNA Array Core Facility (TSRI; La Jolla, CA). The resulting CEL files were processed using the ‘oligo’ R package [20 (link)].

Shannon C.P., Balshaw R., Chen V., Hollander Z., Toma M., McManus B.M., FitzGerald J.M., Sin D.D., Ng R.T, & Tebbutt S.J. (2017). Enumerateblood – an R package to estimate the cellular composition of whole blood from Affymetrix Gene ST gene expression profiles. BMC Genomics, 18, 43.

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Total RNA Extraction from Blood Samples

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Total RNA was extracted in batches at either at the Mercy Hospital for Women (FOX study, corticosteroid and stillbirth cohorts) or the University College London (EVERREST study), using the PAXgene® Blood miRNA Kit (Pre-Analytix, Hombrechtikon, Switzerland) according to the manufacturer’s instructions. In brief, samples were thawed and centrifuged and pellets washed and re-suspended in buffer. Proteins were digested using proteinase K, and cellular debris removed. The supernatant of the resulting flow-through was collected and isopropanol added. Samples were then centrifuged through PAXgene RNA spin columns, and genomic DNA contamination was removed using DNAse digestion. Purified RNA was eluted, and the concentration of RNA was determined by NanoDrop® ND-8000 8-sample spectrophotometer (ThermoFischer Scientific, Waltham, USA).

Hannan N.J., Stock O., Spencer R., Whitehead C., David A.L., Groom K., Petersen S., Henry A., Said J.M., Seeho S., Kane S.C., Gordon L., Beard S., Chindera K., Karegodar S., Hiscock R., Pritchard N., Kaitu’u-Lino T.J., Walker S.P, & Tong S. (2020). Circulating mRNAs are differentially expressed in pregnancies with severe placental insufficiency and at high risk of stillbirth. BMC Medicine, 18, 145.

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Total RNA Extraction from Whole Blood

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We extracted total RNA from whole blood samples using the PAXgene Blood miRNA Kit (PreAnalytix, Qiagen, Hilden, Germany), following the manufacturer’s protocol. We quantified the isolated RNA using a ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA) and removed DNA from the samples using RNase-free DNase (Promega, Madison, WI, USA), also according to the manufacturer’s instructions. We checked the integrity of the RNA with an Agilent 2100 Bioanalyzer, in conjunction with the RNA 6000 Nano kit (Agilent Technologies, CA, USA). RNA integrity number (RIN) value of the samples was of 7.0 or higher.

Gómez-Garre P., Periñán M.T., Jesús S., Bacalini M.G., Garagnani P., Mollenhauer B., Pirazzini C., Provini F., Trenkwalder C., Franceschi C, & Mir P. (2022). Transcriptomic analysis reveals an association of FCGBP with Parkinson’s disease. NPJ Parkinson's Disease, 8, 157.

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Profiling Peripheral Blood RNA Using Illumina Microarrays

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We used PAXgene blood RNA tubes (PreAnalytiX GmbH, Hombrechtikon, Switzerland) as per the kit instructions for the collection of peripheral blood samples (2 × 2.5 ml) in both the sessions. Further, we used PAXgene blood miRNA Kit (PreAnalytiX GmbH, Hombrechtikon, Switzerland) as per the kit instructions for the isolation of total RNA. Next, we tested the purified RNA samples for purity and concentration using the NanoDrop 1000 v.3.7 (Thermo Fisher Scientific, USA). In addition, we used the Ambion's Human GLOBINclear^TM kit (Applied Biosystems, USA) as per the kit insert, for the depletion of globin mRNA. Further, we used the 2100 Bioanalyzer (Agilent Technologies, Germany) to measure the RNA integrity of the samples, before diluting to 50 ng/μl using RNase-free water. A total of 2 μg of RNA was assayed on the Illumina HumanHT-12 v4 bead array (Illumina Inc.; San Diego, CA, USA), which targets more than 47,000 probes.

Kanduri C., Kuusi T., Ahvenainen M., Philips A.K., Lähdesmäki H, & Järvelä I. (2015). The effect of music performance on the transcriptome of professional musicians. Scientific Reports, 5, 9506.

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Peripheral Blood RNA Isolation

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Peripheral blood (2.5 ml) was collected from each participant using BD® Vacutainer Safety-Lok™ Blood Collection Set and drawn into PAXgene® Blood RNA tube (PreAnalytiX). Purification of total RNA including miRNA was conducted using; PAXgene® Blood miRNA Kit (PreAnalytiX) with silica-membrane technology. The quantity of total RNA was measured by NanoDrop® ND-1000 spectrophotometer. Meanwhile, the quality of total RNA and small RNA were analysed by the 2100 Bioanalyzer (Agilent) using Eukaryote Total RNA Nano assay (Agilent) for RNA Integrity Number (RIN) and Small RNA assay (Agilent) for the amount (%) of miRNA.

Mohd Yacob A., Muhamad N.A., Chang K.M., Akmal Hisham H., Mat Yusoff Y, & Ibrahim L. (2022). Hsa-miR-181a-5p, hsa-miR-182-5p, and hsa-miR-26a-5p as potential biomarkers for BCR-ABL1 among adult chronic myeloid leukemia treated with tyrosine kinase inhibitors at the molecular response. BMC Cancer, 22, 332.

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RNA Extraction from Whole Blood

Cited 1 time

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Samples preparation, DNA extraction, library preparation and quality control are detailed in Malvisi et al. (2016) [8 (link)]. In brief, samples used were taken during obligatory routine animal sanitary controls by an authorised veterinarian. No ethical approval was required, in compliance with the European Directive 2010/63/UE and the Italian Regulation D. Lgs n. 26/2014. Whole blood was collected into PAXgene^® Blood RNA tubes (PreAnalytiX GmbH). Total RNA was extracted using the PAXgene^® Blood miRNA kit (PreAnalytiX GmbH). RNA concentration was measured by a NanoDrop^™ 1000 spectrophotometer (Thermo Scientific) and RNA integrity was assessed with an Agilent 2200 TapeStation system (Agilent Technologies).

Malvisi M., Curti N., Remondini D., De Iorio M.G., Palazzo F., Gandini G., Vitali S., Polli M., Williams J.L, & Minozzi G. (2020). Combinatorial Discriminant Analysis Applied to RNAseq Data Reveals a Set of 10 Transcripts as Signatures of Exposure of Cattle to Mycobacterium avium subsp. paratuberculosis. Animals : an Open Access Journal from MDPI, 10(2), 253.

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Quantifying GSTM1 and GSTM5 Expression

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RNA was isolated from peripheral blood leukocytes collected with PAXgene Blood RNA tubes (PreAnalytiX GmbH, Switzerland) per manufacturer’s instructions. Tubes were stored at room temperature for 6 hours and then stored at −80C until RNA isolation (within two weeks after specimen collection). RNA was isolated with the Qiagen QIAcube (Qiagen, Valencia, CA) and PAXgene Blood miRNA Kit (PreAnalytiX GmbH, Switzerland) per manufacturer’s instruction. cDNA was synthesized with reverse transcription reagents (TaqMan; Applied Biosystems, Darmstadt, Germany) according to the manufacturer’s protocol. The final cDNA concentration was determined by nanodrop and diluted to 50ng/μL.
Gene expression assays (TaqMan; Applied Biosystems) were obtained and used for PCR analysis. Probes used were GSTM1 (hs01683722_gh) and GSTM5 (hs00757076_m1). Eukaryotic 18S rRNA (Hs99999901_s1) served as an internal control because of its constant expression level across the studied sample sets. Real-time RT-PCR (TaqMan; Applied Biosystems) was performed (ABI Prism 7900 Sequence Detection System; Applied Biosystems) using the ΔΔC_T method, which provides normalized expression values. The amount of target mRNA was compared among the groups of interest. All reactions were performed in technical triplicates.

Hunter AA I.I.I., Smit-McBride Z., Anderson R., Bordbari M.H., Ying G.S., Kim E.S., Park S.S., Telander D.G., Dunaief J.L., Hjelmeland L.M, & Morse L.S. (2015). GSTM1 and GSTM5 Genetic Polymorphisms and Expression in Age-related Macular Degeneration. Current eye research, 41(3), 410-416.

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Comprehensive RNA Extraction from Whole Blood

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Samples used were taken during obligatory routine animal sanitary controls by an authorized veterinarian. No ethical approval was required, in compliance with the European Directive 2010/63/UE and the Italian regulation D. Lgs n. 26/2014. Whole blood was collected into PAXgene^® Blood RNA Tubes (PreAnalytiX GmbH) and kept at room temperature for at least two hours before freezing at -20°C for 24 hours before being transferred at -80°C until processing, as recommended by the manufacturer. Total RNA was extracted using the PAXgene^® Blood miRNA Kit (PreAnalytiX GmbH) according to the manufacturer’s protocol. Total RNA was eluted in a final volume of 80 μL. RNA concentration was measured by NanoDrop^™ 1000 spectrophotometer (Thermo Scientific) and RNA integrity was assessed with an Agilent 2200 TapeStation system (Agilent Technologies).

Malvisi M., Palazzo F., Morandi N., Lazzari B., Williams J.L., Pagnacco G, & Minozzi G. (2016). Responses of Bovine Innate Immunity to Mycobacterium avium subsp. paratuberculosis Infection Revealed by Changes in Gene Expression and Levels of MicroRNA. PLoS ONE, 11(10), e0164461.

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Transcriptomic analysis of bovine tuberculosis

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Every 3 months, whole blood was collected from HD, LD and control calves in PAXgene® Blood RNA system tubes (PreAnalytix GmbH, Hombrechtikon, Switzerland). Total RNA was extracted from whole blood using the PAXgene® blood miRNA kit (PreAnalytix GmbH) as per kit protocol. The quality of total RNA was determined using an RNA integrity number (Agilent RNA 6000 NanoChip on 2100 Bioanalyzer, Agilent Technologies, Santa Clara, CA, USA). Moreover, 5–10 μg of extracted total RNA was processed using RNeasy Plus Micro kit (Qiagen, Mississauga, ON, Canada) to remove any genomic DNA carryover. The RNA was quantified using NanoDrop ND-1000 (NanoDrop Technologies, Wilmington, DE, USA). Although total RNA was extracted from whole blood collected from trial calves at 3, 6, 9, 12 and 15 months, only total RNA from 6 and 9 months after MAP exposure was used for gene expression analysis by microarray.

David J., Barkema H.W., Guan L.L, & De Buck J. (2014). Gene-expression profiling of calves 6 and 9 months after inoculation with Mycobacterium avium subspecies paratuberculosis. Veterinary Research, 45(1), 96.

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