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Evaluation of optimal RNA extraction method from human carotid atherosclerotic plaque.

Authors :
Ahmed S
Shaffer A
Geddes T
Studzinski D
Mitton K
Pruetz B
Long G
Shanley C
Source :
Cardiovascular pathology : the official journal of the Society for Cardiovascular Pathology [Cardiovasc Pathol] 2015 May-Jun; Vol. 24 (3), pp. 187-90. Date of Electronic Publication: 2014 Dec 05.
Publication Year :
2015

Abstract

Investigating molecular mechanisms involved in the formation of carotid atherosclerotic plaques has been challenging. Isolating high-quality RNA from plaque tissue can be difficult because of acellularity, calcification, and degradation. It is essential that the mRNA isolated from this tissue preserves and reflects the actual relative gene expression. Two common methods for RNA preservation, snap-freezing and stabilizing reagent, were compared using surgically resected human carotid atherosclerotic tissue. In addition, isolation methods were compared for integrity and quantity: column-based extraction, phenol-based extraction, and a combination of the two. We found that using a stabilizing reagent with column filtration resulted in the lowest yield and quality. Phenol-based extraction resulted in higher yields but also increased fragmentation. Snap-frozen tissue coupled with column-based extraction yielded the highest quality. The higher quality and quantity RNA obtained when processing snap-frozen tissue with column-based extraction make it possible to use difficult sample types for molecular downstream applications.<br /> (Copyright © 2014 Elsevier Inc. All rights reserved.)

Details

Language :
English
ISSN :
1879-1336
Volume :
24
Issue :
3
Database :
MEDLINE
Journal :
Cardiovascular pathology : the official journal of the Society for Cardiovascular Pathology
Publication Type :
Academic Journal
Accession number :
25534148
Full Text :
https://doi.org/10.1016/j.carpath.2014.11.005