Evaluation of Ribonucleic Acid (RNA) amplification efficiency to determine optimal RNA concentration ranges from formalin-fixed paraffin-embedded rodent tissues for gene expression analysis

被引：0
|
作者：
Euler, Catherine [1 ,2 ]
Isaacs, Michael [3 ]
Simic, Damir [4 ]
Van Vleet, Terry [1 ,2 ]
Gillham, Susan [1 ,2 ]
Peden, W. Michael [1 ,2 ]
机构：
[1] Bristol Myers Squibb, Dept Pathol, Vernon, IN 47620 USA
[2] Bristol Myers Squibb, Dept Toxicol, Vernon, IN USA
[3] Indiana Univ Sch Med, Indianapolis, IN 46202 USA
[4] Johnson & Johnson, Spring House, PA USA
来源：
JOURNAL OF HISTOTECHNOLOGY | 2015年 / 38卷 / 01期
关键词：
Formalin-fixed paraffin-embedded (FFPE); Polymerase chain reaction (PCR) efficiency; Real-time polymerase chain reaction (RT-qPCR); Ribonucleic acid (RNA) integrity; RNA integrity; MESSENGER-RNA; SAMPLES;
D O I：
10.1179/2046023614Y.0000000051
中图分类号：
Q2 [细胞生物学];
学科分类号：
071009 ; 090102 ;
摘要：
The goal of this study was to establish Ribonucleic Acid (RNA) amplification efficiencies and determine optimal concentration ranges for downstream real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analyses with RNA isolated from formalin-fixed paraffin-embedded (FFPE) and frozen rat tissues. MessengerRNAwas isolated using magnetic particle processing technology. RNAconcentrations and integrity were assessed from frozen and FFPE rat liver, lung, kidney, heart, mammary gland, pancreas, skin, and duodenum, and cycle thresholds (Cts) were obtained from each dilution of an RNA template. PCR amplification efficiencies and optimal template ranges were calculated for FFPE and frozen tissue samples. Calculated percent efficiency evaluations for FFPE and frozen tissues showed that FFPE samples, at the RNA concentrations tested, were of acceptable quality for downstream gene expression analyses.
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页码：2 / 8
页数：7
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