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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Main Authors: 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
Format: Article
Language:English
Published: BMC 2020-03-01
Series:BMC Research Notes
Subjects:
Online Access:http://link.springer.com/article/10.1186/s13104-020-04980-z
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spelling 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
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