A Protocol for Extraction of High-quality RNA and DNA from Peanut Plant Tissues |
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Authors: | Dongmei Yin Haiying Liu Xingguo Zhang Dangqun Cui |
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Affiliation: | (1) Henan Agricultural University, 450002 Zhengzhou, China;(2) Henan Normal University, 453007 Xinxiang, China |
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Abstract: | Peanuts are an increasingly important global food source. However, until recently the lack of effective protocols for the extraction of nucleic acids has made molecular studies of peanut development and maturation difficult. Here, we describe a method to isolate high-quality RNA and DNA from peanut tissue and have successfully applied this method to peanut plant roots, stems, leaves, flowers, and seeds. Spectrophotometric analysis showed that the average yields of total RNA from 100 mg of peanut materials ranged from 24.52 to 74.6 μg, and those of genomic DNA from the same tissues ranged from 23.47 to 57.68 μg. Using this protocol, we obtained OD260/280 values between 1.9 and 2.0 and isolated RNA which could be reverse transcribed in a manner suitable for RT-qPCR and expression analysis. In addition, genomic DNA isolated using this method produced reliable restriction enzyme digestion patterns and could be used for Southern blot hybridization. |
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