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Methods for the in vitro - dna - amplification, for genomic cloning and for genetic mapping.

机译:体外-dna-扩增,基因组克隆和遗传作图的方法。

摘要

A method for cloning genomic DNA comprises the steps of: (a) digesting genomic DNA with a restriction enzyme that generates multiple different cohesive-end sequences; (b) contacting the digested genomic DNA with a linker, said linker comprising a reporter nucleotide sequence and a cohesive-end sequence complementary to a cohesive-end sequence generated by the restriction enzyme of step (a) under conditions such that a linker is joined to a restriction fragment of genomic DNA; and (c) amplifying the product of step (b). Likewise, a method for mapping genomic DNA restriction fragments comprises the steps of: (a) digesting genomic DNA with a restriction enzyme that generates multiple different cohesive-end sequences; (b) amplifying the restriction fragment products of step (a); (c) determining the cohesive-end sequence of each of said restriction fragments; and in accordance therewith, (d) aligning the restriction fragments in a physical map.
机译:克隆基因组DNA的方法包括以下步骤:(a)用限制性酶消化基因组DNA,所述限制性酶产生多个不同的内聚末端序列; (b)使消化的基因组DNA与接头接触,所述接头包含报告核苷酸序列和与由步骤(a)的限制酶产生的内切末端序列互补的内切末端序列,在连接子被连接的条件下对基因组DNA的限制性片段; (c)扩增步骤(b)的产物。同样,用于定位基因组DNA限制片段的方法包括以下步骤:(a)用产生多个不同内聚末端序列的限制酶消化基因组DNA; (b)扩增步骤(a)的限制性片段产物; (c)确定每个所述限制性片段的内聚末端序列;据此,(d)在物理图中比对限制性片段。

著录项

  • 公开/公告号DE69011101T2

    专利类型

  • 公开/公告日1995-01-26

    原文格式PDF

  • 申请/专利权人 GENENTECH INC US;

    申请/专利号DE1990611101T

  • 发明设计人 MILLER HARVEY US;

    申请日1990-02-15

  • 分类号C12N15/10;C12Q1/68;

  • 国家 DE

  • 入库时间 2022-08-22 04:08:23

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