Exponential megapriming PCR (EMP) cloning--seamless DNA insertion into any target plasmid without sequence constraints.
basic_science · Level V
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- Record sourced from PubMed, PMID 23300917.
- Also identified by DOI 10.1371/journal.pone.0053360 and PMC identifier PMC318348.
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Abstract
We present a fast, reliable and inexpensive restriction-free cloning method for seamless DNA insertion into any plasmid without sequence limitation. Exponential megapriming PCR (EMP) cloning requires two consecutive PCR steps and can be carried out in one day. We show that EMP cloning has a higher efficiency than restriction-free (RF) cloning, especially for long inserts above 2.5 kb. EMP further enables simultaneous cloning of multiple inserts.