The expression data of RA were downloaded from Gene Expression Omnibus (GEO) DataStets (https://www.ncbi.nlm.nih.gov/sites/GDSbrowser?acc=GDS3794 ) and corresponding reference [21 (link)]. In the study, a complete genome-wide transcript profiling of peripheral blood mononuclear cells (PBMCs) from 18 RA patients and 15 age- and sex-matched healthy controls were collected. The information about the demographical and clinical characteristics of RA patients and controls was presented in Table S1 . The total RNA was isolated from PBMCs using the PAXgene RNA isolation kit (PreAnalytix). The concentration and profiles of total RNA were determined using a NanoDrop ND-1000 (Wilmington) and a Bioanalyzer 2100 (Agilent). Two hundred nanograms of RNA was reverse transcribed. The cDNA was transcribed and cRNA was labeled with biotin-16-UTP. Labeled probe hybridization to Illumina BeadChips human-6v2 was carried out using Illumina’s BeadChip 6v2 protocol. Beadchips were scanned on the Illumina BeadArray 500GX Reader. The preliminary data analysis were performed and normalized by Illumina BeadStudio software. Then the real-time PCR was further performed to detect mRNA expression. Differential analysis per gene was performed with one-way analysis of variance (ANOVA) and P values were adjusted to control the false discovery rate (FDR, 5%).
Paxgene rna isolation kit
Manufactured by Preanalytix
Sourced in Switzerland
The PAXgene RNA isolation kit is a laboratory product designed to extract and purify RNA from blood samples. It is used to prepare RNA for various downstream analysis applications.
Automatically generated - may contain errors
2 protocols using paxgene rna isolation kit
1
Transcriptomic Profiling of RA Patients
2
Profiling PBMC Transcriptome in Rheumatoid Arthritis
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The gene expression profile of PBMC of RA was downloaded from the GEO database (access number: GSE15573). Briefly, peripheral blood specimens were collected from 18 RA patients and 15 healthy controls, matched for age and gender. All were Caucasians living in Paris, France. Total RNA was isolated from PBMC using the PAXgene RNA isolation kit (PreAnalytix, Hombrechtikon, Switzerland). The isolated total RNA was transcribed into cRNA and labelled biotin-16-UTP. Illumina human-6 v2.0 expression beadchip (Illumina, San Diego, California) (containing 12 600 oligonucleotide probes) was applied for microarray hybridisation following standard protocol. Hybridisation signals were scanned by Illumina BeadArray 500GX Reader and analysed by Illumina BeadScan software 2.3.0.13 (Illumina). BeadStudio software (version 1.5.0.34) was used for preliminary data analysis and quality control. The generated microarray data were expressed as log2 ratios of the fluorescence intensities of RA samples divided by the fluorescence intensities of control samples. Normalisation was conducted using the "normalise quantiles" function in the BeadStudio software. A detailed description of study subjects, experimental procedures and analysis approaches can be found in the original study.5 (link)
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