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CN102115742A 一种植物中总核酸的同步提取方法 无效 - 驳回

一种植物中总核酸的同步提取方法
摘要：
本发明属于生物技术领域，公开了一种植物中总核酸的同步提取方法。将生物材料粉碎后与CTAB裂解液混合，水浴45-60min，加入与CTAB裂解液等体积的由氯仿和异戊醇组成的混合试剂，混匀；离心，取上清，加入等体积的由水饱和酚、氯仿及异戊醇组成的混合试剂，混匀；离心，取上清，加入1/30体积的NaAc和1/10体积的无水乙醇，混匀，在冰上静置5min后，再加入等体积的由氯仿和异戊醇组成的混合试剂；离心，取上清，加入1/10体积的NaAc和两倍体积的无水乙醇，-20℃静置1h；离心，弃上清，用70％的乙醇洗涤沉淀1～5次，干燥得到包含基因组DNA和总RNA的总核酸。该发明同步提取烟草总核酸，对于今后开展基因克隆、表达分析等一系列分子生物学研究具有非常重要的意义。
摘要（英）：

The invention belongs to the technical field of biology, and discloses a synchronous extraction method of total nucleic acid in plant. The method comprises the steps of: smashing a biological material, mixing the smashed biological material with CTAB (cetyltrimethyl ammonium bromide) lysis solution, performing the water bath for 45-60 min, adding the mixed reagent which has the same volume as that of the CTAB lysis solution and consists of chloroform and isoamyl alcohol, and evenly mixing; centrifuging, talking supernate, adding the isometric mixed reagent which consists of phenol werer, the chloroform and the isoamyl alcohol, and evenly mixing; centrifuging, talking the supernate, adding NaAc with 1/30 volume and absolute ethyl alcohol with 1/10 volume, evenly mixing, stewing for 5min on ice, and adding the isometric mixed reagent which consists of the chloroform and the isoamyl alcohol; centrifuging, talking the supernate, adding the NaAc with 1/30 volume and the absolute ethyl alcohol with doubled volume, and stewing at -20 DEG C for 1h; and centrifuging, removing the supernate, depositing by 70% alcohol in a washing way for 1-5 times, and drying to obtain the total nucleic acid which contains genome DNA (deoxyribose nucleic acid) and total RNA (rbose nucleic acid). The synchronous extraction method of the tobacco total nucleic acid has a very important meaning to a series of molecular biology research such as the gene clone, expression, analysis and the like.

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