Publication Date:
2012-11-04
Description:
RNA snap ™ is a simple and novel method that recovers all intracellular RNA quantitatively (〉99%), faster (〈15 min) and less expensively (~3 cents/sample) than any of the currently available RNA isolation methods. In fact, none of the bacterial RNA isolation methods, including the commercial kits, are effective in recovering all species of intracellular RNAs (76–5700 nt) with equal efficiency, which can lead to biased results in genome-wide studies involving microarray or RNAseq analysis. The RNA snap ™ procedure yields ~60 µg of RNA from 10 8 Escherichia coli cells that can be used directly for northern analysis without any further purification. Based on a comparative analysis of specific transcripts ranging in size from 76 to 5700 nt, the RNA snap ™ method provided the most accurate measure of the relative amounts of the various intracellular RNAs. Furthermore, the RNA snap ™ RNA was successfully used in enzymatic reactions such as RNA ligation, reverse transcription, primer extension and reverse transcriptase–polymerase chain reaction, following sodium acetate/ethanol precipitation. The RNA snap ™ method can be used to isolate RNA from a wide range of Gram-negative and Gram-positive bacteria as well as yeast.
Keywords:
RNA characterisation and manipulation
Print ISSN:
0305-1048
Electronic ISSN:
1362-4962
Topics:
Biology
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