新疆野苹果果实总RNA提取方法的筛选

doi:10.13866/j.azr.2019.02.20

Abstract

Abstract: Malus sieversii fruits are rich in polysaccharides,polyphenols,soluble pectin and other secondary metabolites. Therefore,it is relatively difficult to extract the total RNA from M.sieversii fruits. In order to choose an effective extraction method,five RNA extraction methods including the TaKaRa Kit,Trans Kit,improved CTAB method by Tian Wei,modified CTAB method I and CTAB method Ⅱwere compared using the fruits at five different developmental stages. Results showed that neither kits method was able to be used to extract the RNA from M.sieversii fruits. The quality of total RNA extracted with three modified CTAB methods was good,with two distinct electrophoresis bands of 28S and 18S rRNA,the OD₂₆₀/OD₂₈₀ values ranged from 1.8 to 2.2,and the OD₂₆₀/OD₂₃₀ value was higher than 2.0. But only the modified CTAB method I was an effective method to yield high quantity of total RNA. The average extraction values of fruits at five different developmental stages were ranged from 116.79 μg·g^-1 to 474.76 μg·g^-1. For the modified CTAB method I,the impurity in the RNA was effectively removed by extending LiCl precipitation time and placing extraction under 4 ℃ for overnight aging (not longer than 16 hours)instead of -20 ℃ for 15 min. Although this method took 2 days,the method was easy to operate,and the extraction cost was low. Therefore,the modified CTAB method I is an effective method for total RNA extraction from M.sieversii fruits (especially for the ripe fruits),which could be used in further molecular biology research.

Key words: Malus sieversii Roeml, total RNA, extraction method, improved CTAB method

XIA Chun-lan, WEN Zhi-bin, ZHANG Yu-fang. Choosing of Methods for Total RNA Extraction from Malus sieversii Fruits in Xinjiang[J].Arid Zone Research, 2019, 36(2): 437-443.

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Choosing of Methods for Total RNA Extraction from Malus sieversii Fruits in Xinjiang

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