COMPARISON OF RNA EXTRACTION METHODS FROM SPONGES
Abstract
Many sponges produce a variety of chemical compounds that inhibit the downstream applications of both DNA and RNA. Therefore, robust extraction methods need to be developed. Three extraction methods were compared; TRIzol® reagent, TRIzol® plus columns from the FastDNA® Spin kit for soil and lastly the FastRNA® Green kit. We investigated these methods in terms of yield of RNA recovered and quality of RNA. Presence of inhibitory compounds was ascertained by the ability to produce double stranded cDNA. Results indicate that all methods were able to extract stable RNA that could be converted to cDNA. However, the amount of RNA and 260/280 ratios varied between the methods, while the size of RNA was similar for all three methods. Yields of cDNA varied from 900ng to 500ng. In conclusion the FastRNA® Green kit produced not only largest amount of RNA per gram of sponge but also the highest quality based on conversion to cDNA.Downloads
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2018-05-23
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