The paraffin-embedded RNA metric (PERM) for RNA isolated from formalin-fixed, paraffin-embedded tissue

被引:16
|
作者
Chung, Joon-Yong [1 ]
Cho, Hanbyoul [1 ,2 ]
Hewitt, Stephen M. [1 ]
机构
[1] NCI, Expt Pathol Lab, Pathol Lab, Ctr Canc Res,NIH, MSC 1500, Bethesda, MD 20892 USA
[2] Yonsei Univ, Coll Med, Gangnam Severance Hosp, Dept Obstet & Gynecol, Seoul, South Korea
关键词
formalin-fixed; paraffin-embedded; RNA integrity; molecular pathology; TIME RT-PCR; QUALITY ASSESSMENT; DNA ASSAY; QUANTIFICATION; INTEGRITY; RECOVERY; IMPACT;
D O I
10.2144/000114415
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
RNA isolated from formalin-fixed, paraffin-embedded (FFPE) tissue is commonly evaluated in both investigative and diagnostic pathology. However, the quality of the data is directly impacted by RNA quality. The RNA integrity number (RIN), an algorithm based on a combination of electrophoretic features, is widely applied to RNA isolated from paraffin-embedded tissue, but it is a poor indicator of the quality of that RNA. Here we describe the novel paraffin-embedded RNA metric (PERM) for quantifying the quality of RNA from FFPE tissue. The PERM is based on a formula that approximates a weighted area-under-the-curve analysis of an electropherogram of the extracted RNA. Using biochemically degraded RNAs prepared from experimentally fixed mouse kidney specimens, we demonstrate that PERM values correlate with mRNA transcript measurements determined using the QuantiGene system. Furthermore, PERM values correlate with real-time PCR data. Our results demonstrate that the PERM can be used to qualify RNA for different end-point studies and may be a valuable tool for molecular studies using RNA extracted from FFPE tissue.
引用
收藏
页码:239 / +
页数:5
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