RNA recovery and detection of mRNA by RT-PCR from preserved prokaryotic samples.

Abstract:

:The effectiveness of maintaining prokaryotic RNA in Synechococcus and Pseudomonas cells, fixed in 96% ethanol, 4% paraformaldehyde, or suspended in RNAlater, and held in cold storage for 3 months was compared. Fluorometric determination of the RNA extracted from Synechococcus and Pseudomonas cells indicated that the cell storage treatments tested were equally effective at maintaining their total RNA content. There was not any detectable decrease in the quantity of RNA isolated from the preserved samples during storage. Intact mRNA transcripts of the RuBisCO (rbcL) and nir genes were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) from preserved bacterial cells throughout 3 months of storage. In contrast, RT-PCR failed to amplify the mRNA of the rbcL and nitrite reductase genes in unfixed and/or unpreserved bacterial samples, suggesting that bacterial mRNA can be well maintained during a prolonged storage when cells are preserved properly. In addition, RNAlater is a useful reagent for the storage and maintenance of high quality RNA in unfrozen samples.

journal_name

FEMS Microbiol Lett

authors

Bachoon DS,Chen F,Hodson RE

doi

10.1111/j.1574-6968.2001.tb10745.x

keywords:

subject

Has Abstract

pub_date

2001-07-24 00:00:00

pages

127-32

issue

2

eissn

0378-1097

issn

1574-6968

pii

S0378109701002518

journal_volume

201

pub_type

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