A specialized blood collection device, cell-free RNA BCT (RNA-BCT) is investigated for its ability to stabilize RNA in intact blood cells at room temperature post-phlebotomy. |
The RT-qPCR-based analysis is carried out to study the expression levels of GAPDH, c-fos and p53 mRNA of neutrophils isolated from blood samples collected in K3EDTA and RNA-BCT devices. |
Results indicate that expression levels of these mRNAs are stable in neutrophils obtained from RNA-BCT blood for up to 3 days at room temperature as compared to the neutrophils obtained from K3EDTA blood. |
1 Introduction
2 Materials and Methods
2.1 Recruitment of Blood Donors
2.2 Blood Collection
2.3 Isolation of Neutrophils from Whole Blood Using Magnetic Beads
2.4 Total Cellular RNA Isolation
2.5 RNA Yield and Purity
2.6 RT-qPCR
2.7 Statistical Analysis
3 Results
3.1 RNA Yield and Purity
Sample type | Total RNA yield (µg) | 260/280 nm ratio | 260/230 nm ratio |
---|---|---|---|
Mean ± SD (p) | Mean ± SD (p) | Mean ± SD (p) | |
K3EDTA day 0 | 3.7 ± 0.7 | 1.9 ± 0.09 | 1.8 ± 0.51 |
K3EDTA day 1 | 3.5 ± 1 (0.77) | 1.9 ± 0.06 (1) | 1.6 ± 0.27 (0.25) |
K3EDTA day 2 | 2.9 ± 1.4 (0.32) | 1.8 ± 0.06 (0.74) | 1.6 ± 0.15 (0.63) |
K3EDTA day 3 | 3.6 ± 0.9 (0.81) | 1.8 ± 0.11 (0.9) | 1.7 ± 0.77 (0.84) |
RNA-BCT day 0 | 2.9 ± 0.5 (0.06) | 1.8 ± 0.1 (0.64) | 1.32 ± 0.4 (0.26) |
RNA-BCT day 1 | 3.4 ± 0.7 (0.58) | 1.8 ± 0.05 (0.80) | 1.5 ± 0.3 (0.35) |
RNA-BCT day 2 | 4.3 ± 2 (0.58) | 1.8 ± 0.09 (0.43) | 1.5 ± 0.3 (0.50) |
RNA-BCT day 3 | 3.7 ± 1 (0.91) | 1.7 ± 0.3 (0.43) | 1.7 ± 0.1 (0.69) |