Mathew Rebecca, Toufiq Mohammed, Mattei Valentina, Al Hashmi Muna, Shobha Manjunath Harshitha, Syed Ahamed Kabeer Basirudeen, Calzone Rita, Cugno Chiara, Chaussabel Damien, Deola Sara, Tomei Sara
Omics Core, Research Branch, Out Patient Clinic, Sidra Medicine, PO 26999, Doha, Qatar.
System Biology, Research Branch, Out Patient Clinic, Sidra Medicine, Doha, Qatar.
BMC Res Notes. 2020 Mar 13;13(1):150. doi: 10.1186/s13104-020-04980-z.
Transcriptome analysis of human whole blood is used to discover biomarkers of diseases and to assess phenotypic traits. Here we have collected small volumes of blood in Tempus solution and tested whether different storage conditions have an impact on transcriptomic profiling. Fifty µl of blood were collected in 100µl of Tempus solutions, freezed at - 20 °C for 1 day and eventually thawed, stored and processed under five different conditions: (i) - 20 °C for 1 week; (ii) +4 °C for 1 week; (iii) room temperature for 1 week; (iv) room temperature for 1 day, - 20 °C for 1 day, room temperature until testing at day 7, (v) - 20 °C for 1 week, RNA was isolated and stored in GenTegra solution. We used 272 immune transcript specific assays to test the expression profiling using qPCR based Fluidigm BioMark HD dynamic array.
RNA yield ranged between 0.17 and 1.39µg. Except for one sample, RIN values were > 7. Using Principal Component Analysis, we saw that the storage conditions did not drive sample distribution. The condition that showed larger variability was the RT-FR-RT (room temperature-freezing-room temperature), suggesting that freezing-thawing cycles may have a worse effect on data reproducibility than keeping the samples at room temperature.
对人类全血进行转录组分析以发现疾病生物标志物并评估表型特征。在此，我们收集了少量置于Tempus溶液中的血液，并测试了不同储存条件是否会对转录组分析产生影响。将50微升血液收集到100微升Tempus溶液中，在-20°C下冷冻1天，最终解冻，并在五种不同条件下储存和处理：(i)-20°C保存1周；(ii)+4°C保存1周；(iii)室温保存1周；(iv)室温保存1天，-20°C保存1天，直至第7天测试前一直保持室温；(v)-20°C保存1周，分离RNA并储存在GenTegra溶液中。我们使用272种免疫转录物特异性检测方法，基于Fluidigm BioMark HD动态阵列的qPCR检测表达谱。
RNA产量在0.17至1.39微克之间。除一个样本外，RNA完整性数（RIN）值均>7。通过主成分分析，我们发现储存条件并未驱动样本分布。变异性较大的条件是室温-冷冻-室温（RT-FR-RT），这表明冻融循环对数据重现性的影响可能比将样本保存在室温下更严重。
