Linker tailing: unphosphorylated linker oligonucleotides for joining DNA termini.
Linker tailing: unphosphorylated linker oligonucleotides for joining DNA termini.
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连接尾部：用于连接 DNA 末端的未磷酸化连接寡核苷酸。
DOI：
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发表时间：
1984
期刊：
影响因子：
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通讯作者：
J. Lecocq
中科院分区：
文献类型：
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作者：
R. Lathe;M. Kieny;S. Skory;J. Lecocq
目前用于在DNA末端之间插入接头寡核苷酸的程序涉及末端添加接头双链体，然后进行限制性酶切割和再连接。这些方法的缺点是含有内部识别位点的靶DNA分子在接头切割步骤中被切割。我们描述了一种在DNA末端添加接头寡核苷酸的方法，该方法消除了后续限制性内切酶处理的要求。将非磷酸化的接头双链体单独连接到一条链上的靶DNA末端。去除未连接的链在每个末端产生单链突起。两个这样的加尾末端现在可以通过使它们的单链互补尾部退火而连接，从而产生含有单个接头插入的杂合DNA分子。该程序的每一步都进行了研究，以优化重组分子的回收，我们提出了如何应用该方法的例子。
Current procedures for inserting linker oligonucleotides between DNA termini involve terminal addition of linker duplexes followed by restriction enzyme cleavage and religation . Such methods have the disadvantage that target DNA molecules containing internal recognition sites are cut during the linker cleavage step. We describe a method for adding linker oligonucleotides to DNA termini which eliminates the requirement for subsequent restriction enzyme treatment. Nonphosphorylated linker duplexes are ligated to target DNA termini on one strand alone. Removal of the unligated strand generates a single-stranded protrusion at each terminus. Two such tailed termini may now be linked by annealing their single-stranded complementary tails, resulting in a hybrid DNA molecule containing a single linker insertion. Each step of the procedure has been studied to optimize the recovery of recombinant molecules, and we present examples of how the method may be applied.
由于末端磷酸盐的去除，聚脱氧核糖核苷酸在高分辨率聚丙烯酰胺凝胶中的迁移率发生改变。
影响因子：
14.9
作者：
Tapper,DP;Clayton,DA
通讯作者：
Clayton,DA
影响因子：
14.9
作者：
HOOPES, BC;MCCLURE, WR
通讯作者：
MCCLURE, WR