Influence of storage conditions of small volumes of blood on immune transcriptomic profiles
Abstract Objective 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. Fift...
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doaj-f1c1351f255d41c7b37deb5c81ac676b2020-11-25T02:56:33ZengBMCBMC Research Notes1756-05002020-03-011311710.1186/s13104-020-04980-zInfluence of storage conditions of small volumes of blood on immune transcriptomic profilesRebecca Mathew0Mohammed Toufiq1Valentina Mattei2Muna Al Hashmi3Harshitha Shobha Manjunath4Basirudeen Syed Ahamed Kabeer5Rita Calzone6Chiara Cugno7Damien Chaussabel8Sara Deola9Sara Tomei10Omics Core, Research Branch, Out Patient Clinic, Sidra MedicineSystem Biology, Research Branch, Out Patient Clinic, Sidra MedicineOmics Core, Research Branch, Out Patient Clinic, Sidra MedicineOmics Core, Research Branch, Out Patient Clinic, Sidra MedicineOmics Core, Research Branch, Out Patient Clinic, Sidra MedicineSystem Biology, Research Branch, Out Patient Clinic, Sidra MedicineAdvanced Cell Therapy Core, Sidra MedicineAdvanced Cell Therapy Core, Sidra MedicineSystem Biology, Research Branch, Out Patient Clinic, Sidra MedicineAdvanced Cell Therapy Core, Sidra MedicineOmics Core, Research Branch, Out Patient Clinic, Sidra MedicineAbstract Objective 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. Results 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.http://link.springer.com/article/10.1186/s13104-020-04980-zStorageBloodTrancriptomeQPCRImmune profilingTempus |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Rebecca Mathew Mohammed Toufiq Valentina Mattei Muna Al Hashmi Harshitha Shobha Manjunath Basirudeen Syed Ahamed Kabeer Rita Calzone Chiara Cugno Damien Chaussabel Sara Deola Sara Tomei |
spellingShingle |
Rebecca Mathew Mohammed Toufiq Valentina Mattei Muna Al Hashmi Harshitha Shobha Manjunath Basirudeen Syed Ahamed Kabeer Rita Calzone Chiara Cugno Damien Chaussabel Sara Deola Sara Tomei Influence of storage conditions of small volumes of blood on immune transcriptomic profiles BMC Research Notes Storage Blood Trancriptome QPCR Immune profiling Tempus |
author_facet |
Rebecca Mathew Mohammed Toufiq Valentina Mattei Muna Al Hashmi Harshitha Shobha Manjunath Basirudeen Syed Ahamed Kabeer Rita Calzone Chiara Cugno Damien Chaussabel Sara Deola Sara Tomei |
author_sort |
Rebecca Mathew |
title |
Influence of storage conditions of small volumes of blood on immune transcriptomic profiles |
title_short |
Influence of storage conditions of small volumes of blood on immune transcriptomic profiles |
title_full |
Influence of storage conditions of small volumes of blood on immune transcriptomic profiles |
title_fullStr |
Influence of storage conditions of small volumes of blood on immune transcriptomic profiles |
title_full_unstemmed |
Influence of storage conditions of small volumes of blood on immune transcriptomic profiles |
title_sort |
influence of storage conditions of small volumes of blood on immune transcriptomic profiles |
publisher |
BMC |
series |
BMC Research Notes |
issn |
1756-0500 |
publishDate |
2020-03-01 |
description |
Abstract Objective 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. Results 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. |
topic |
Storage Blood Trancriptome QPCR Immune profiling Tempus |
url |
http://link.springer.com/article/10.1186/s13104-020-04980-z |
work_keys_str_mv |
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