The focus on sample quality: Influence of colon tissue collection on reliability of qPCR data
Autor: | Mikael Kubista, Jana Slyskova, Vendula Novosadova, Vaclav Liska, Paolo Verderio, Jens Björkman, Ludmila Vodickova, Lucie Langerova, Karel Veskrna, Ondrej Vycital, Vlasta Korenkova, Miroslav Levy, Pavel Vodicka, Sara Pizzamiglio |
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Jazyk: | angličtina |
Rok vydání: | 2016 |
Předmět: |
0301 basic medicine
Quality Control Tissue Fixation Colon Nitrogen Organ Preservation Solutions Preservation Biological Computational biology Biology Bioinformatics Real-Time Polymerase Chain Reaction DNA Ribosomal Article Specimen Handling 03 medical and health sciences 0302 clinical medicine High-Throughput Screening Assays RNA Ribosomal 18S Humans RNA Neoplasm Cryopreservation Multidisciplinary Gene Expression Profiling Carcinoma RNA Snap freezing Reproducibility of Results DNA Neoplasm Ribosomal RNA Neoplasm Proteins Gene expression profiling Gene Expression Regulation Neoplastic 030104 developmental biology Real-time polymerase chain reaction DNA Topoisomerases Type I 030220 oncology & carcinogenesis Colonic Neoplasms Nucleic acid |
Zdroj: | Scientific Reports |
ISSN: | 2045-2322 |
DOI: | 10.1038/srep29023 |
Popis: | 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. |
Databáze: | OpenAIRE |
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