Back to Search Start Over

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

Authors :
Nevin Belder
Öznur Coşkun
Beyza Doğanay Erdoğan
Berna Savaş
Arzu Ensari
Hilal Özdağ
Source :
Genomics Data, Vol 7, Iss , Pp 303-306 (2016)
Publication Year :
2016
Publisher :
Elsevier, 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. Keywords: Microarray, Gene expression, FFPE, Optimization

Subjects

Subjects :
Genetics
QH426-470

Details

Language :
English
ISSN :
22135960
Volume :
7
Issue :
303-306
Database :
Directory of Open Access Journals
Journal :
Genomics Data
Publication Type :
Academic Journal
Accession number :
edsdoj.4ede028ca048407bb072fa7120c15545
Document Type :
article
Full Text :
https://doi.org/10.1016/j.gdata.2016.02.006