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| Determination of the Structure of m3G Cap RNA by Co-immunoprecipitation and Microarray |
| JIA Dong1, ZHANG Bin2, MA Rui-yan1, TANG Gui-gang1, ZHAO Jie1, WANG Wen-hua1, ZHOU Lin1 |
1. College of Agriculture, Shanxi Agricultural University, Taigu 030801, China;
2. College of Horticulture, Shanxi Agricultural University, Taigu 030801, China |
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Abstract Objective:To establish a protocol to determine the structure of RNA 5'end cap by using co-immunoprecipitation(Co-IP) in conjunction with microarray technology. Methods: Total RNAs isolated from Caenorhabditis elegans were precipitated with the antibody against M3G cap structure. For Microarray analysis, the probes were designed for 127 non-coding RNAs (ncRNAs) of C.elegans. The cap structure was determined and RNA transcription was also analyzed. Results: 37 ncRNAs was isolated and identified as containing m3G cap structure. This number is similar with other species, for which m3G cap RNAs have been found. Conclusions: It is feasible and effective to determine the structure of ncRNA 5' end cap by using Co-IP followed by Microarray analysis. This method is of great specificity and high sensitivity, which maybe applied to the large-scale RNA cap structure identification, the discovery of novel RNA and the identification of RNA functions.
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Received: 04 November 2013
Published: 25 February 2014
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