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. 2009 Dec 22;4:201–209. doi: 10.4137/bmi.s3774

Figure 1. Validation of assay condition.

Figure 1

A) Blood storage at 4 °C. B) Applicability of frozen whole blood. Heparinized human blood samples were collected from 6 adult donors (APEX Research Institute), and were stored at 4 °C for 4–6 (Day 0), 24 (Day 1), or 48 hours (Day 2). Blood samples were then stimulated with 10, 1, 0.1 and 0 Gy of radiation (cesium-137) in triplicate, followed by the incubation at 37 °C for an additional 2 hours. One set of samples was stored at −80 °C, and the other set was processed immediately without freezing procedure. Frozen samples were analyzed 1 week later. ACTB (•), CDKN1A (○) and BBC3 (▴) mRNA were quantified as described in the Methods, and fold increase was calculated using the values of unexposed samples with 2 hours incubation. Blood volume used for each analysis was 150 μl (50 μl in triplicate), and each gene was amplified individually by TaqMan (CDKN1A and BBC3) or SYBR green (ACTB), respectively. Symbols are the mean value from each subject.