Rapid Assembly of DNA via Ligase Cycling Reaction (LCR)

Rapid Assembly of DNA via Ligase Cycling Reaction (LCR)
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DOI:
10.1007/978-1-4939-6343-0_8
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发表时间:
2017-01-01
期刊:
SYNTHETIC DNA: METHODS AND PROTOCOLS
影响因子:
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通讯作者:
Chandran, Sunil
Chandran, Sunil
中科院分区:
其他
文献类型:
--
作者:
Chandran, Sunil

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将多个DNA部分组装成一个更大的DNA结构是大多数合成生物学实验室的要求。在这里,我们描述了一种利用连接酶链式反应(LCR)高效、高通量地组装DNA的方法。LCR方法利用在桥联寡核苷酸的指导下连接在一起的不重叠的DNA部分。使用这种方法,我们已经成功地在一次反应中组装了多达20个DNA部分,或构建了大小达26kb的DNA。
The assembly of multiple DNA parts into a larger DNA construct is a requirement in most synthetic biology laboratories. Here we describe a method for the efficient, high-throughput, assembly of DNA utilizing the ligase chain reaction (LCR). The LCR method utilizes non-overlapping DNA parts that are ligated together with the guidance of bridging oligos. Using this method, we have successfully assembled up to 20 DNA parts in a single reaction or DNA constructs up to 26 kb in size.