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Room temperature stabilization of gene transcripts and genomic DNA in whole blood
Author(s) -
Steven P,
Stassinopoulos Angela,
Whitney Scott,
Ohgi Sharron,
Clement Omo,
MullerCohn Judy,
Muller Rolf
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb310
Subject(s) - genomic dna , whole blood , genotyping , dna , gene , gene expression , rna , biology , real time polymerase chain reaction , reference genes , computational biology , microbiology and biotechnology , genetics , genotype , immunology
Genomic and mitochondrial DNA and gene expression profiles from blood samples are increasingly being used to diagnose specific diseases, monitor disease progression and assess patient responses to medical treatment. However, such applications require reliable preservation of total RNA and DNA in patient samples during collection, transport and storage. Numerous agents threaten DNA and RNA profiles in collected blood samples, including nucleolytic attack, oxidative damage and hydrolysis. Such damage can severely disrupt genotyping and DNA sequencing efforts. Transcription profiles are highly dynamic and can change rapidly during and after blood collection using current methods, potentially affecting interpretation of the expression analysis. In this study, we compare DNAgard Blood, PAXgene Blood DNA and freezer storage in their capacities to preserve genomic DNA integrity in human whole blood for 6 months. In a separate analysis, we compare RNA stabilization properties of PAXgene Blood RNA, freezer storage and a novel stabilization formulation by assessing changes in the relative expression of a panel of genes in whole blood specimens over the course of 7 days using qRT‐PCR and the Human Common Cytokines PCR Array (SABiosciences). Our results demonstrate the importance of selecting an appropriate blood storage method to obtain reliable gene expression and genomic data.