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

(2020) PLOS ONE. 15(2).
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Abstract
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., (NucliSENS) 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.
Keywords
General Biochemistry, Genetics and Molecular Biology, General Agricultural and Biological Sciences, General Medicine, HIGH-QUALITY RNA, MESSENGER-RNA, EXPRESSION, AMPLIFICATION, OPTIMIZATION, CLOSTRIDIUM, COLLECTION, SYSTEM, STEP, DNA

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MLA
Rodriguez Sánchez, Antonio, et al. “Comparison of Procedures for RNA-Extraction from Peripheral Blood Mononuclear Cells.” PLOS ONE, vol. 15, no. 2, 2020, doi:10.1371/journal.pone.0229423.
APA
Rodriguez Sánchez, A., Duyvejonck, H., Van Belleghem, J., Gryp, T., Van Simaey, L., Vermeulen, S., … Vaneechoutte, M. (2020). Comparison of procedures for RNA-extraction from peripheral blood mononuclear cells. PLOS ONE, 15(2). https://doi.org/10.1371/journal.pone.0229423
Chicago author-date
Rodriguez Sánchez, Antonio, Hans Duyvejonck, Jonas Van Belleghem, Tessa Gryp, Leen Van Simaey, Stefan Vermeulen, Els Van Mechelen, and Mario Vaneechoutte. 2020. “Comparison of Procedures for RNA-Extraction from Peripheral Blood Mononuclear Cells.” PLOS ONE 15 (2). https://doi.org/10.1371/journal.pone.0229423.
Chicago author-date (all authors)
Rodriguez Sánchez, Antonio, Hans Duyvejonck, Jonas Van Belleghem, Tessa Gryp, Leen Van Simaey, Stefan Vermeulen, Els Van Mechelen, and Mario Vaneechoutte. 2020. “Comparison of Procedures for RNA-Extraction from Peripheral Blood Mononuclear Cells.” PLOS ONE 15 (2). doi:10.1371/journal.pone.0229423.
Vancouver
1.
Rodriguez Sánchez A, Duyvejonck H, Van Belleghem J, Gryp T, Van Simaey L, Vermeulen S, et al. Comparison of procedures for RNA-extraction from peripheral blood mononuclear cells. PLOS ONE. 2020;15(2).
IEEE
[1]
A. Rodriguez Sánchez et al., “Comparison of procedures for RNA-extraction from peripheral blood mononuclear cells,” PLOS ONE, vol. 15, no. 2, 2020.
@article{8677182,
  abstract     = {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., (NucliSENS) 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.},
  articleno    = {e0229423},
  author       = {Rodriguez Sánchez, Antonio and Duyvejonck, Hans and Van Belleghem, Jonas and Gryp, Tessa and Van Simaey, Leen and Vermeulen, Stefan and Van Mechelen, Els and Vaneechoutte, Mario},
  issn         = {1932-6203},
  journal      = {PLOS ONE},
  keywords     = {General Biochemistry,Genetics and Molecular Biology,General Agricultural and Biological Sciences,General Medicine,HIGH-QUALITY RNA,MESSENGER-RNA,EXPRESSION,AMPLIFICATION,OPTIMIZATION,CLOSTRIDIUM,COLLECTION,SYSTEM,STEP,DNA},
  language     = {eng},
  number       = {2},
  pages        = {17},
  title        = {Comparison of procedures for RNA-extraction from peripheral blood mononuclear cells},
  url          = {http://dx.doi.org/10.1371/journal.pone.0229423},
  volume       = {15},
  year         = {2020},
}

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