Traditional cut-paste DNA cloning is often limited by the availability of restriction enzyme sites. Here, we described the complementary annealing mediated by exonuclease (CAME), in which the insert or vector fragment is amplified to carry sequences complementary to the other, and both fragments are modified by exonuleases to create directional single-stranded overhangs. The two recessed DNA fragments are joined through complementary strand annealing. The CAME is highly efficient for cloning the DNA of at least 12 kb and single DNA fragment out of a complex DNA sample. Moreover, the application of CAME greatly improved the efficiency of site-directed mutagenesis.

中文翻译：

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外切酶介导的互补退火：一种无缝克隆和调节定点诱变的方法

传统的剪切粘贴 DNA 克隆通常受到限制性内切酶位点可用性的限制。在这里，我们描述了由外切核酸酶 (CAME) 介导的互补退火，其中插入或载体片段被扩增以携带与另一个互补的序列，并且两个片段都被外切核酸酶修饰以产生定向单链突出端。两个凹陷的 DNA 片段通过互补链退火连接。CAME 对于从复杂的 DNA 样本中克隆至少 12 kb 的 DNA 和单个 DNA 片段非常有效。此外，CAME的应用大大提高了定点诱变的效率。