| 一种快速高效提取酵母质粒的方法
| A Fast, Efficient and Economical Method for Isolating Plasmid From Yeast
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| DOI：10.3969/j.issn.1673-1689.2013.11.012
| 中文关键词: 酵母菌 质粒提取 玻璃珠液氮法
| 英文关键词: yeast plasmid extraction glass beads-liquid nitrogen method
| 基金项目:国家自然科学基金项目(31071807,31272168);周口师范学院实验室开放项目(K201337)
| 作者
| 单位
| 张怡
| 周口师范学院植物遗传与分子育种重点实验室,河南周口,466001
| 刘坤
| 周口师范学院植物遗传与分子育种重点实验室,河南周口,466001
| 曹鹏
| 周口师范学院植物遗传与分子育种重点实验室,河南周口,466001
| 魏森
| 周口师范学院植物遗传与分子育种重点实验室,河南周口,466001
| 韦丹丹
| 周口师范学院植物遗传与分子育种重点实验室,河南周口,466001
| 杜建
| 周口师范学院植物遗传与分子育种重点实验室,河南周口,466001
| 李鹏坤
| 周口师范学院植物遗传与分子育种重点实验室,河南周口,466001
| 李成伟
| 周口师范学院植物遗传与分子育种重点实验室,河南周口,466001
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| 中文摘要:
| 质粒提取是遗传工程操作的重要步骤。酵母细胞壁坚韧,不易破壁,使酵母质粒提取变得非常困难。为了获得一种快速、简便、高效、稳定的提取酵母质粒的方法,利用琼脂糖凝胶电泳、核酸蛋白测定仪和PCR扩增等方法,对玻璃珠液氮法、蜗牛酶法和试剂盒法提取的酵母质粒,在产率、纯度和对后续实验的影响等方面进行了比较。结果表明,与另外两种方法比较,玻璃珠液氮法提取酵母质粒具有效率高、质量好、成本低、时间短、操作简单的优点,可作为实验室规模化提取酵母质粒的常用方法。
| 英文摘要:
| Plasmid extraction is an important but difficult step of the genetic engineering operations,because of the yeasts cells with tough and unbreakable cell wall. For getting one fast,simple,efficient and stable method of plasmid extraction from yeasts,we employed the techniques of agarose gel electrophoresis,spectrophotometer and PCR amplification,and compared the yield,purity of plasmids which were isolated from yeasts with the methods of glass beads-liquid nitrogen,snailase and kit. The results showed that the method of glass beads-liquid nitrogen had the advantages of high efficiency,good quality,low cost,simple operation in contrast to the other two methods,and it can be used as a laboratory scale commonly method for extraction of yeast plasmid.
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