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Sample GSM1574933 Query DataSets for GSM1574933
Status Public on Dec 31, 2015
Title NSCLC 25
Sample type RNA
 
Source name plasma
Organism Homo sapiens
Characteristics gender: Male
age at interview: 56
smoking status: Ever smoker
morphology (for patients only): 8071/3
disease state: Non-small cell lung cancer patient
Treatment protocol Peripheral blood was collected in EDTA tubes at the time of interview.
Growth protocol n/a
Extracted molecule total RNA
Extraction protocol Total RNA was isolated from 300L of plasma using NucleoSpin® miRNA Plasma kit (Macherey-Nagel, Düren, Germany) according to the manufacturer’s protocol with Proteinase K digest, addition of 2μg of glycogen carrier and DNAse digest steps. All samples were spiked-in with 10pmol of Arabidopsis thaliana synthetic miR-159a (synthesized by Eurofins MWG Operon, Ebersberg, Germany) to control for variations in the RNA preparation step.
Label FAM
Label protocol Expression levels of 754 miRNAs (Sanger miRBase v14) were quantified using the TaqMan Human MicroRNA Array A + B Card Set v3.0 (Applied Biosystems, Foster City, CA) as per the manufacturer's instructions (including pre-amplification).
 
Hybridization protocol n/a
Scan protocol n/a
Description Case
Data processing The data were analysed in HTqPCR package using R Bioconductor. miRNAs with undetermined Ct values in more than 120 samples were filtered out. Data were quantile normalized. Following normalization Ct values with an interquartile range (IQR) of less than 1.5 and endogenous controls were removed from subsequent analysis, and limma analysis was performed to identify differentially regulated miRNA between cases and controls. With limma, a one-factorial linear model is fitted for each miRNA and the standard errors (SE) are moderated using an empirical Bayes model resulting in moderated t-statistics for each miRNA. These analysis were performed separately for CARD A and CARD B.
Matrix normalized worksheet reports normalized signal (quantile normalization). Fold Change worksheet reports fold changes across all cases and controls.
 
Submission date Dec 30, 2014
Last update date Dec 31, 2015
Contact name Magdalena B Wozniak
Organization name International Agency for Research on Cancer
Department Genetic Epidemiology
Street address 150 cours Albert Thomas
City Lyon
ZIP/Postal code 69008
Country France
 
Platform ID GPL18942
Series (1)
GSE64591 Circulating microRNAs as non-invasive biomarkers for early detection of non-small-cell lung cancer

Supplementary data files not provided
Processed data are available on Series record

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