Successful molecular analyses of human solid tissues require intact biological material with well-preserved nucleic acids, proteins, and other cell structures. Pre-analytical handling, comprising of the collection of material at the operating theatre, is among the first critical steps that influence sample quality. The aim of this study was to compare the experimental outcomes obtained from samples collected and stored by the conventional means of snap freezing and by PAXgene Tissue System (Qiagen). These approaches were evaluated by measuring rRNA and mRNA integrity of the samples (RNA Quality Indicator and Differential Amplification Method) and by gene expression profiling. The collection procedures of the biological material were implemented in two hospitals during colon cancer surgery in order to identify the impact of the collection method on the experimental outcome. Our study shows that the pre-analytical sample handling has a significant effect on the quality of RNA and on the variability of qPCR data. PAXgene collection mode proved to be more easily implemented in the operating room and moreover the quality of RNA obtained from human colon tissues by this method is superior to the one obtained by snap freezing.

Biomedical Centre Medical School Pilsen Charles University Prague Pilsen Czech Republic

Deparment of Surgery Teaching Hospital and Medical School Pilsen Charles University Prague Pilsen Czech Republic

Institute of Biology and Medical Genetics 1st Faculty of Medicine Charles University Prague Prague Czech Republic

Institute of Biotechnology BIOCEV Centre Czech Academy of Sciences Průmyslová 595 252 42 Vestec u Prahy Czech Republic

Institute of Experimental Medicine Czech Academy of Sciences Prague Czech Republic

Surgical Department Thomayer Hospital 1st Faculty of Medicine Charles University Prague Prague Czech Republic

TATAA Biocenter AB Göteborg Sweden

Unit of Medical Statistics Biometry and Bioinformatics Fondazione Istituto di Ricovero e Cura a Carattere Scientifico Istituto Nazionale dei Tumori Milan Italy

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