Microarray analysis was performed on three independent MEFs/genotype using Illumina MouseWG-6 v2.0 Expression BeadChip (Department of Pathology, Cambridge University). Normalised Log2 values were determined and average Log Fold Change (LogFC) calculated for each comparison. Pathway analysis of genes differentially expressed (>1.3 fold) between genotypes was performed using Ingenuity IPA analysis software (ingenuity.com ) and statistical significance (P<0.05) of canonical pathways determined by Benjamini-Hochberg multiple testing correction. Relative gene expression was depicted by heatmaps generated using GENE-E software and statistical significance (P<0.05) determined by t-test. Gene expression changes were validated by qPCR using ROCHE Universal Probe Library System or Life Technologies probes and all data normalised to 18S expression.
Universal probe library system
Manufactured by Thermo Fisher Scientific
The Universal Probe Library System is a qPCR (quantitative Polymerase Chain Reaction) platform designed for gene expression analysis. It provides a comprehensive set of pre-designed probes that can be used for a wide range of target genes, enabling efficient and accurate measurement of gene expression levels.
Automatically generated - may contain errors
Lab products found in correlation
Mousewg 6 v2.0 expression beadchip, by Illumina (2 mentions)
Viia 7 real time pcr system, by Thermo Fisher Scientific (2 mentions)
Geneglobe data analysis center, by Qiagen (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Dnase 1, by Thermo Fisher Scientific (1 mentions)
Dneasy blood and tissue kit, by Qiagen (1 mentions)
Tissuelyser, by Qiagen (1 mentions)
Nanodrop nd 100, by Thermo Fisher Scientific (1 mentions)
Rt2 profiler pcr array, by Qiagen (1 mentions)
High pure rna isolation kit, by Roche (1 mentions)
4 protocols using universal probe library system
1
Transcriptome Analysis of Genetically Modified MEFs
2
Transcriptome Analysis of Genetically Modified MEFs
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Microarray analysis was performed on three independent MEFs/genotype using Illumina MouseWG-6 v2.0 Expression BeadChip (Department of Pathology, Cambridge University). Normalised Log2 values were determined and average Log Fold Change (LogFC) calculated for each comparison. Pathway analysis of genes differentially expressed (>1.3 fold) between genotypes was performed using Ingenuity IPA analysis software (ingenuity.com ) and statistical significance (P<0.05) of canonical pathways determined by Benjamini-Hochberg multiple testing correction. Relative gene expression was depicted by heatmaps generated using GENE-E software and statistical significance (P<0.05) determined by t-test. Gene expression changes were validated by qPCR using ROCHE Universal Probe Library System or Life Technologies probes and all data normalised to 18S expression.
3
Autophagy Gene Expression in Cold Exposure
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Tissue samples were homogenized in TRIzol reagent (Life Technologies, 15596-026) with a TissueLyser (Qiagen). Total RNA was extracted, treated with DNase I (Life Technologies, 18068015), and reverse transcribed to complementary DNA using the M-MuLV Reverse Transcription Kit (NEB). DNA is extracted by using DNeasy Blood and Tissue kit (Qiagen 69504). qPCR was performed with the TaqMan method (Roche Universal ProbeLibrary System) on a ViiA 7 Real-Time PCR System (Applied Biosystems). Relative expression was calculated using the ΔΔCt method with normalization to β-actin. All the experiments were repeated by at least 3 times by using independent samples. Autophagy PCR array was from Qiagen (PAMM-084Z) and beta-actin was used as control gene. The data were analyzed by GeneGlobe Data Analysis Center (Qiagen). RNA samples were from 3 cold-challenged mice and 4 mice from thermoneutrality.
4
Mouse Macrophage RNA Expression Profiling
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RNA was extracted from mouse peritoneal macrophage by Roche High Pure RNA Isolation Kit (Roche). RNA quality was assessed by the Nanodrop ND-100 (Nanodrop Technologies, Inc., Wilmington DE). The CDNA was reverse transcribed by Iscript™ II. qPCR was performed with the TaqMan method (Roche Universal ProbeLibrary System) on a ViiA 7 Real-Time PCR System (Applied Biosystems). Relative expression was calculated using the ΔΔCt method with normalization to β-actin. All the experiments were repeated by at least 3 times by using independent samples. Mouse NFkB signaling pathway gene expression profiling was assessed with RT2 Profiler PCR Array (Qiagen, PAMM-025ZC-2) and β actin was used as control gene. The data were analyzed by GeneGlobe Data Analysis Center (Qiagen).
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