Preanalytical aspects of quantitative TaqMan real-time RT-PCR: Applications for TF and VEGF mRNA quantification

Isaksson, Helena S.; Nilsson, Torbjörn

doi:10.1016/j.clinbiochem.2006.02.003

Cited by 9 publications

(2 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Similarly, the EDTA-anticoagulated blood has been successfully used to measure the production of cytokines at protein and RNA level upon stimulation with tetanus (38,39). The study of Isaksson et al (40) demonstrated that different laboratory tube types did not have significant influence on VEGF mRNA stability. This finding may be explained by different processing delays.…”

Section: Discussionmentioning

confidence: 99%

Influence of Pre-analytical Variables on VEGF Gene Expression and Circulating Protein Concentrations

Azimi‐Nezhad

Lambert²,

Ottone³

et al. 2012

Biopreservation and Biobanking

View full text Add to dashboard Cite

The most reliable conditions for measuring both circulating VEGF and its gene expression are to reduce time between blood collection and centrifugation, and to avoid multiple freeze-thaw cycles. Serum collection tubes with no additive and no separator were less sensitive to the pre-analytical variations analyzed in this study. Freezing delay had a significant influence on VEGF protein expression in tissue samples.

show abstract

Section: Discussionmentioning

confidence: 99%

Influence of Pre-analytical Variables on VEGF Gene Expression and Circulating Protein Concentrations

Azimi‐Nezhad

Lambert²,

Ottone³

et al. 2012

Biopreservation and Biobanking

View full text Add to dashboard Cite

show abstract

“…For some genes, however, the reference method resulted in similar gene expression levels, so the use of a stabilization reagent does not seem to be essential for every gene. This has also been shown for the quantification of tissue factor (F3) and vascular endothelial growth factor, with comparable results derived from stabilized and unstabilized samples (28 ). Therefore, each researcher must carefully evaluate the preanalytical handling of the samples and choose a system for RNA ex vivo stabilization in case of gene expression analysis in later project phases with unknown targets at start of study.…”

Section: Discussionmentioning

confidence: 99%

Preanalytical mRNA Stabilization of Whole Bone Marrow Samples

et al. 2007

View full text Add to dashboard Cite

Background: Gene expression profiling is a useful tool for cancer diagnosis and basic research. A major limitation is that, even during short-term storage of native specimens of peripheral blood or bone marrow (BM) and/or RNA isolation, significant changes of gene expression pattern can occur because of gene induction, repression, and RNA degradation. Methods: We investigated the effectiveness of a newly developed RNA stabilization and preparation system for BM specimens (PAXgene™ Bone Marrow RNA System) over time. We analyzed 256 RNA samples, processed from 64 BM specimens. Results: Although the overall RNA yield (normalized to 1 ؋ 10 7 leukocytes) was not different, the RNA preparation using unstabilized reference samples had an ϳ3 times higher failure rate. With the PAXgene system, we observed significantly higher RNA integrity compared with the reference RNA preparation system (P <0.01). In the stabilized samples, we found very high pairwise correlation in gene expression (⌬⌬C T 0.16 -0.53) for the analyzed genes (GATA1, RUNX1, NCAM1, and SPI1) after 48-h storage compared with immediate preparation of RNA (2 h after BM collection). However, we found major differences in half of the analyzed genes using the reference RNA isolation procedure (⌬⌬C T 1.07 and 1.32). Conclusions: The PAXgene system is able to stabilize RNA from clinical BM samples and is suitable to isolate high-quality and -quantity RNA.

show abstract

Specimen Collection, Handling, and Processing

Hengesbach

Gerlach

2012

Modern Clinical Molecular Techniques

View full text Add to dashboard Cite

Preanalytical aspects of quantitative TaqMan real-time RT-PCR: Applications for TF and VEGF mRNA quantification

Cited by 9 publications

References 18 publications

Influence of Pre-analytical Variables on VEGF Gene Expression and Circulating Protein Concentrations

Influence of Pre-analytical Variables on VEGF Gene Expression and Circulating Protein Concentrations

Preanalytical mRNA Stabilization of Whole Bone Marrow Samples

Specimen Collection, Handling, and Processing

Contact Info

Product

Resources

About