Comparison of Six Different Pretreatment Methods for Blood RNA Extraction

Liu, Xiaopan; Li, Qiyuan; Wang, Xian; Zhou, Xiaolin; Liao, Qiuyan; He, Xuheng; Zhang, Jiuling; Sun, Jianbo; Wu, Jang‐Yen; Cheng, Lei; Yong, Zhang

doi:10.1089/bio.2014.0090

Cited by 17 publications

(17 citation statements)

References 12 publications

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“…Therefore, the way samples are stored preceding RNA extraction as well as the kit of choice to extract RNA are crucial. Thus far, only a limited number of studies compared storage and extraction of RNA from human samples in a methodological manner [19,23,24]. In this study, different storage media (RNL, QZL or cognate lysis buffer (respectively EML, Table 3.…”

Section: Discussionmentioning

confidence: 99%

Comparison of procedures for RNA-extraction from peripheral blood mononuclear cells

et al. 2020

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RNA quality and quantity are important factors for ensuring the accuracy of gene expression analysis and other RNA-based downstream applications. Thus far, only a limited number of methodological studies have compared sample storage and RNA extraction procedures for human cells. We compared three commercially available RNA extraction kits, i.e., (Nucli-SENS) easyMAG, RNeasy (Mini Kit) and RiboPure (RNA Purification Kit-blood). In addition, additional conditions, such as storage medium and storage temperature of human peripheral blood mononuclear cells were evaluated, i.e., 4˚C for RNAlater or -80˚C for QIAzol and for the respective cognate lysis buffers; easyMAG, RNeasy or RiboPure. RNA was extracted from aliquots that had been stored for one day (Run 1) or 83 days (Run 2). After DNase treatment, quantity and quality of RNA were assessed by means of a NanoDrop spectrophotometer, 2100 Bioanalyzer and RT-qPCR for the ACTB reference gene. We observed that high-quality RNA can be obtained using RNeasy and RiboPure, regardless of the storage medium, whereas samples stored in RNAlater resulted in the least amount of RNA extracted. In addition, RiboPure combined with storage of samples in its cognate lysis buffer yielded twice as much RNA as all other procedures. These results were supported by RT-qPCR and by the reproducibility observed for two independent extraction runs.PLOS ONE | https://doi.org/10.1371/journal.pone.

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Section: Discussionmentioning

confidence: 99%

Comparison of procedures for RNA-extraction from peripheral blood mononuclear cells

et al. 2020

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“…However, RNA is susceptible to numerous factors such as the time of sample collection, transportation conditions, protectants used, pre-treatments, and extraction techniques prior to transcriptome profiling. These could affect the quality of RNA isolated and, eventually, the results of the analysis [ 26 ]. Given that transcriptome profiles from whole blood offer huge sources of information during infections, it is important to employ this approach especially when dealing with systemic infections such as LF, likewise in localized skin infections such as onchocerciasis with several infection phenotypes.…”

Section: Prospects Of Whole Blood Microarray In Filarial Infectionmentioning

confidence: 99%

The Potentials and Pitfalls of Microarrays in Neglected Tropical Diseases: A Focus on Human Filarial Infections

Kwarteng

Ahuno

2016

Microarrays

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Data obtained from expression microarrays enables deeper understanding of the molecular signatures of infectious diseases. It provides rapid and accurate information on how infections affect the clustering of gene expression profiles, pathways and networks that are transcriptionally active during various infection states compared to conventional diagnostic methods, which primarily focus on single genes or proteins. Thus, microarray technologies offer advantages in understanding host-parasite interactions associated with filarial infections. More importantly, the use of these technologies can aid diagnostics and helps translate current genomic research into effective treatment and interventions for filarial infections. Studying immune responses via microarray following infection can yield insight into genetic pathways and networks that can have a profound influence on the development of anti-parasitic vaccines.

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“…Several studies targeting the quanti cation of gene expression pro les have been performed to better understand human and veterinary diseases [1][2][3][4][5][6][7][8]. In this context, blood specimens are the most common applied matrix since peripheral blood may be considered less invasive and valuable source of RNA in vertebrates [9]. However, RNA blood is highly vulnerable to degradation, which may consequently in uence on the results of RNA studies to varying level [9].…”

Section: Introductionmentioning

confidence: 99%

“…In this context, blood specimens are the most common applied matrix since peripheral blood may be considered less invasive and valuable source of RNA in vertebrates [9]. However, RNA blood is highly vulnerable to degradation, which may consequently in uence on the results of RNA studies to varying level [9].…”

Section: Introductionmentioning

confidence: 99%

How long does the mRNA remains stable in untreated whole bovine blood?

Giglioti

Azevedo

Oliveira

et al. 2021

Preprint

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Background: High quality and quantity of messenger RNA (mRNA) are required for accuracy of gene expression studies and other RNA-based downstream applications. Since RNA is considered a labile macromolecular prone to degradation, which may result in falsely altered gene expression patterns, several commercial stabilizing reagents have been developed aiming to keep RNA stable for long period. However, for studies involving large number of experimental samples, the high costs related to these specific reagents may constitute a barrier. Methods and Results: In this context the present study was designed aiming to evaluate the stability of mRNA in whole bovine blood collected in EDTA tubes during storage at common fridge (4°C). Whole blood samples were collected from six Holstein calves and submitted to RNA extraction in each different interval: immediately after blood sampling (< 2 h), at 1-day post-sampling (dps), 2 dps, 3 dps, 7 dps and 14dps intervals. RNA integrity and purity were evaluated, and RT-qPCR assays were run using seven different genes (B2M, ACTB, PPIA, GAPDH, YWHAZ, CD4 and IFN-γ) aiming to evaluate the presence of altered gene transcription during storage. All extracted RNA samples presented high purity, while optimal integrity and unaltered gene expression were observed in whole experimental group up to 3 days of storage.Conclusion: Bovine blood RNA remained stable in K3EDTA tubes for 3 days stored at common fridge and can be successfully and accurately used for gene expression studies.

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Comparison of Six Different Pretreatment Methods for Blood RNA Extraction

Cited by 17 publications

References 12 publications

Comparison of procedures for RNA-extraction from peripheral blood mononuclear cells

Comparison of procedures for RNA-extraction from peripheral blood mononuclear cells

The Potentials and Pitfalls of Microarrays in Neglected Tropical Diseases: A Focus on Human Filarial Infections

How long does the mRNA remains stable in untreated whole bovine blood?

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