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microRNA Profiling from Blood Samples

The regulatory effects microRNA of are widespread and microRNAs have recently been implicated in various diseases of the blood, stem cells, brain, and other tissues.

MicroRNAs are expressed in the blood

A study involving sickle cell diseases revealed that there is significant microRNA expression in red blood cells1 and another showed that microRNAs are highly expressed in the ischemic brain and can be detected in blood samples2.

MicroRNA expression analysis of blood samples can further our understanding of diseases of the blood

Analysis of microRNA expression in samples extracted from peripheral blood has shed some light on the causes of systemic lupus erythematosus3 and various aberrantly expressed microRNAs were detected in granulocytes, mononuclear cells, platelets, and reticulocytes of patients with polycythemia vera4.  Altered microRNA expression has also been detected in granulocytes isolated from blood samples of patients with primary myelofibrosis5.

MicroRNA extraction protocol for blood samples

There is a need to extract microRNA from whole blood samples for expression analysis to continue the research effort toward better understanding of these and other diseases.

Two important considerations when isolating RNA from whole blood samples are:  first, the sample collection method as well as the stability of the RNA must be considered.  It is very important to limit the time from sample collection to microRNA analysis.  Next, as with all sample types, it is very important to use a total RNA preparation procedure that does not remove the low molecular weight RNA fractions.

Several published studies6,7 have demonstrated good expression results when the PAXgene™ system is used as a blood sample collection method.  Although these studies made use of the manufacturer’s subsequent recommended RNA extraction procedure (or slight modification thereof) our extensive in-house development work has resulted in a proprietary alternative extraction method which we have used repeatedly to generate robust and reliable expression results.

LC Sciences’ recommended procedure for collecting blood samples for microRNA analysis:

  1. Use PAXgene™ blood collection tubes. (Qiagen/BD)
  2. Collect multiple tubes from each donor and make note of the order of collection.
  3. Keep the samples cold and limit the time to analysis. (follow manufacturer's recommendation for storage times/temps)
  4. Send at least two tubes from each donor. (Do not send the first tube collected from a donor)

LC Sciences can perform the RNA extraction from your samples if you prefer, or perform the RNA extraction yourself using the PAXgene™ recommended procedure or one of the many kits available for extraction of microRNA from blood samples:

Total RNA Isolation Kit - Norgen Biotek

Mouse Ribopure-Blood RNA isolation kit – Ambion

You should expect a yield of about 5-10µg of total RNA from each PAXgene™ tube.  See the ordering page for more specifics on QC of your total RNA sample.

For extraction of microRNA from plasma samples we recommend the Norgen Biotek kit.

  1. Chen SY, Wang Y, Telen MJ, Chi JT. (2008) The genomic analysis of erythrocyte microRNA expression in sickle cell diseases. PLoS ONE. 4;3(6):e2360. [abstract]
  2. Jeyaseelan K, Lim KY, Armugam A. (2008) MicroRNA expression in the blood and brain of rats subjected to transient focal ischemia by middle cerebral artery occlusion. Stroke. 39(3):959-66. [abstract]
  3. Dai Y, Huang YS, Tang M, Lv TY, Hu CX, Tan YH, Xu ZM, Yin YB. (2007) Microarray analysis of microRNA expression in peripheral blood cells of systemic lupus erythematosus patients. Lupus. 16(12):939-46. [abstract]
  4. Bruchova H, Merkerova M, Prchal JT. (2008) Aberrant expression of microRNA in polycythemia vera. Haematologica. [Epub ahead of print] [abstract]
  5. Guglielmelli P, Tozzi L, Pancrazzi A, Bogani C, Antonioli E, Ponziani V, Poli G, Zini R, Ferrari S, Manfredini R, Bosi A, Vannucchi AM; MPD Research Consortium. (2007) MicroRNA expression profile in granulocytes from primary myelofibrosis patients. Exp Hematol. 35(11):1708-18. [abstract]
  6. Kruhøffer M, Dyrskjøt L, Voss T, Lindberg RL, Wyrich R, Thykjaer T, Orntoft TF. (2007) Isolation of microarray-grade total RNA, microRNA, and DNA from a single PAXgene blood RNA tube. J Mol Diagn. 9(4):452-8. [abstract]
  7. Liu J, Walter E, Stenger D, Thach D. (2006) Effects of globin mRNA reduction methods on gene expression profiles from whole blood. J Mol Diagn. 8(5):551-8. [abstract]

 

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