Effects of Postmortem Interval and Preservation Method on RNA Isolated from Field-Preserved Avian Tissues

The condor

Zachary A. Cheviron¹ 

关键词: gene expression;    genetic resources;    mRNA;    natural history collections;    postmortem interval;    transcriptomics;   

DOI  :  10.1525/cond.2011.100201

学科分类：动物科学

来源: Central Ornithology Publication Office

【 摘 要 】

Abstract. Studies of gene expression and transcriptome profiling often require the isolation of high-quality mRNA. Here, we examined how preservation method and the lag between collection and processing (postmortem interval; PMI) affect RNA quality and yield in field-collected samples. We subjected tissue samples of 11 House Sparrows (Passer domesticus) to two preservation treatments (flash freezing in liquid nitrogen versus treatment with Ambion's RNAlater prior to freezing) and eight PMI treatments (intervals ranging from 0 to 360 min). Mean RNA yields were significantly higher for flash-frozen tissues. Total RNA yield, as measured by a UV-vis spectrophotometer (Nanodrop), was not negatively associated with PMI in either preservation treatment, and we were able to extract RNA of quality (28S:18S fluorescence ratio >0.9) sufficient for most gene-expression applications from specimens that were held at ambient temperature for up to 6 hr postmortem. These results demonstrate that sufficient quantities of h...

【 授权许可】

CC BY