数字农科院2.0

一种高效提取棉花细胞核的技术(英文)

文献类型： 中文期刊

作者： 彭仁海;刘方;宋国立;王春英;黎绍惠;张香娣;王玉红;王坤波

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期刊名称：Agricultural Science & Technology

ISSN： 1009-4229

年卷期： 2009 年 03 期

页码： 19-21

摘要： [Objective] The experiment aimed to study an efficient method of Nuclei extraction of cotton and provided technical support for constructing large-insert genomic library and sequencing complete genome. [Method] The cotton cotyledons germinated in dark moisture chamber for one week were chopped with a sharp sterile scalpel in a Petri dish which contained ice-cold nucleus isolation buffer (10 mmol/L MgSO4, 5 mmol/L KCl, 0.5 mmol/L HEPES, 1 mg/ml DTT, 0.25% Triton X-100 and 2% PVP40), then the nuclei were collected after selected through 100, 50 and 30 μm nylon meshes and centrifugation. Meanwhile, the tender leaves and cotyledons with different germination time in dark were treated by grinding method and sharp scalpel method. [Result] The chopping with a sharp scalpel method was very simple and rapid, which did not need grind and mercaptoethanol treatment and the successful extraction rate was 100%.[Conclusion] An efficient method of nuclei extraction of cotton with simple, high efficiency, rapid reaction and poison free were established.

分类号： S562

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