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pmid:26981433

Optimization of gene expression microarray protocol for formalin-fixed paraffin-embedded tissues.

Nevin Belder · Öznur Coşkun · Beyza Doğanay Erdoğan · Berna Savaş · Arzu Ensari · Hilal Özdağ

Genomics data · 2016 doi:10.1016/j.gdata.2016.02.006

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complete

Experiment

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title
Optimization of gene expression microarray protocol for formalin-fixed paraffin-embedded tissues.
doi
10.1016/j.gdata.2016.02.006
pmid
26981433
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Nevin Belder
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Öznur Coşkun
[2]
Beyza Doğanay Erdoğan
[3]
Berna Savaş
[4]
Arzu Ensari
[5]
Hilal Özdağ
journal
Genomics data
year
2016
abstract
Formalin-fixed paraffin-embedded (FFPE) tissue is a widely available clinical specimen for retrospective studies. The possibility of long-term clinical follow-up of FFPE samples makes them a valuable source to evaluate links between molecular and clinical information. Working with FFPE samples in the molecular research area, especially using high-throughput molecular techniques such as microarray gene expression profiling, has come into prominence. Because of the harmful effects of formalin fixation process such as degradation of nucleic acids, cross-linking with proteins, and chemical modifications on DNA and RNA, there are some limitations in gene expression profiling studies using FFPE samples. To date many studies have been conducted to evaluate gene expression profiling using microarrays (Thomas et al., Thomas et al. (2013) [1]; Scicchitano et al., Scicchitano et al. (2006) [2]; Frank et al., Frank et al. (2007) [3]; Fedorowicz et al., Fedorowicz et al. (2009) [4]). However, there is still no generally accepted, efficient and standardized procedure for microarray analysis of FFPE samples. This paper describes the microarray data presented in our recently accepted to be published article showing a standard protocol from deparaffinization of FFPE tissue sections and RNA extraction to microarray gene expression analysis. Here we represent our data in detail, deposited in the gene expression omnibus (GEO) database with the accession number GSE73883. Four combinations of two different cRNA/cDNA preparation and labeling protocols with two different array platforms (Affymetrix Human Genome U133 Plus 2.0 and U133_X3P) were evaluated to determine which combination gives the best percentage of present call. The study presents a dataset for comparative analysis which has a potential in terms of providing a robust protocol for gene expression profiling with FFPE tissue samples.
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This paper describes the microarray data presented in our recently accepted to be published article showing a standard protocol from deparaffinization of FFPE tissue sections and RNA extraction to microarray gene expression analysis.
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Group comparisons were performed using Student's t test and one-way ANOVA.
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Array data files with a .CEL extension were preprocessed for background correction and quantile normalization using Robust multiarray averaging (RMA). Paired t test was performed to identify differentially expressed genes.
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Optimization of gene expression microarray protocol for formalin-fixed paraffin-embedde
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high-throughput molecular techniques such as microarray gene expression profiling, has come into pro
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The possibility of long-term clinical follow-up of FFPE samples makes them a valuable source to evaluate links between molecular and clinical information. Working with FFPE samples in the molecular research area, especially using high-throughput molecular techniques such as microarray gene expression profiling, has come into prominence. Because of the harmful effects of formalin fixation process such as degradation of nucleic acids, cross-linking with proteins, and chemical modifications on DNA and RNA, there are some limitations in gene expression profiling studies using FFPE samples. To date many studies have been conducted to evaluate gene expression profiling using microarrays (Thomas et al., Thomas et al. (2013) [1]; Scicchitano et al., Scicchitano et al. (2006) [2]; Frank et al., Frank et al. (2007) [3]; Fedorowicz et al., Fedorowicz et al. (2009) [4]). However, there is still no generally accepted, efficient and standardized procedure for microarray analysis of FFPE samples. This paper describes the microarray data presented in our recently accepted to be published article showing a standard protocol from deparaffinization of FFPE tissue sections and RNA extraction to microarray gene expression analysis. Here we represent our data in detail, deposited in the gene expression omnibus (GEO) database with the accession number GSE73883. Four combinations of two different cRNA/cDNA preparation and labeling protocols with two different array platforms (Affymetrix Human Genome U133 Plus 2.0 and U133_X3P) were evaluated to determine which combination gives the best percentage of present call. 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Paired t test was performed to identify differentially expressed genes."}, {"asset": "source:pmid:26981433", "locator": "sections[9] \"Statistical data analysis\" paragraph 2", "quote": "The entire microarray data set has been deposited at the Gene Expression Omnibus (GEO) database with Accession Number GSE73883."}, {"asset": "source:pmid:26981433", "locator": "sections[2] \"Direct link to deposited data\" paragraph 2", "quote": "http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE73883"}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "The paper presents a standard FFPE microarray workflow and a comparative gene-expression dataset deposited in GEO under accession GSE73883; statistical comparisons used Student’s t test, one-way ANOVA, and paired t test with RMA normalization.", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:26981433"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. 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