Comparative analysis and innovation of a simple and rapid method for high-quality RNA and DNA extraction of kiwifruit

摘要

class="kwd-title">Method name: RNA and DNA extraction for fleshy fruits class="kwd-title">Keywords: RNA and DNA extraction, RT-qPCR, Kiwifruit, Polyphenols, Polysaccharides class="head no_bottom_margin" id="abs0010title">AbstractRNA and DNA extraction is a requirement for the study of gene expression and has an increasingly important role in genetic studies of all fleshy fruits. RNA and DNA extraction is difficult in kiwifruit due to the significant amount of polysaccharides and polyphenols compounds. So far, no commercial kit has been developed specifically for high-quality RNA and DNA extraction in kiwifruit and the common protocols for RNA extraction have poor yields. This study developed a new protocol for high quality RNA extraction in Actinidia deliciosa. According to the results, the average yield of RNA extraction of fruit and leaf of A. deliciosa was ∼2180.7 ng/μl (∼545.175 μg/g FW) and ∼3424.9 ng/μl (∼856.225 μg/g FW), respectively with A260/A280 between 1.95 to 2.07 and A260/A230 higher than 2 indicating high RNA purity. While the averages yield of RNA extraction using previous methods from kiwifruit and leaf was 23 μg/g FW and 527 μg/g FW, respectively. Also, the average yields of genomic DNA from kiwifruit ranged from 52 to 98 ng/μl with A260/A230 between 0.60 to 1.64 and A260/A280 between 1.40 to 1.48. To our knowledge, this is the first report of a highly efficient and rapid method of RNA and DNA extraction in kiwifruit which can be used for a broad spectrum of the all fleshy fruits.