A quick method to isolate RNA from wheat and other carbohydrate-rich seeds.
A quick method to isolate RNA from wheat and other carbohydrate-rich seeds.
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DOI：
10.1007/bf02773401
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发表时间：
2003-01-01
影响因子：
2.1
通讯作者：
Singh, P.
中科院分区：
文献类型：
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作者：
Gurpreet Singh, Gurpreet Singh;Sanjay Kumar, Sanjay Kumar;Singh, P.
从富含碳水化合物的小麦种子中分离RNA的报道尚未发表。由于碳水化合物的存在，已发表的方案产生少量质量差的RNA。在缓冲液（pH 9，150 mM NaCl，1%肌氨酸）中提取种子确保了最大的RNA溶解度和大多数干扰物质的去除。使用基于盐酸胍的缓冲系统纯化提取的RNA。该方案在3.5小时内从100 mg组织中产生高达148 g RNA。A260/A280比值为1.85表明RNA纯度。分离的RNA适合于下游应用，如差异显示。开发的方法被扩展到其他碳水化合物丰富的种子，如大麦和玉米，并取得了成功。
No reports on isolating RNA from carbohydrate-rich wheat seeds have been published. Because of the presence of carbohydrates, published protocols yield small amounts of poor quality RNA. Extracting seeds in a buffer (pH 9, 150 mM NaCl, 1% sarcosyl) ensured maximum RNA solubility and the removal of most interfering substances. Extracted RNA was purified using a guanidine hydrochloride-based buffer system. This protocol yields up to 148 g of RNA from 100 mg of tissue in 3.5 h. An A260/A280 ratio of 1.85 indicates RNA purity. Isolated RNA was amenable to downstream applications such as differential display. The developed method was extended to other carbohydrate-rich seeds, such as barley and maize, with success.