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  4. Exponential Megapriming PCR (EMP) Cloning-Seamless DNA Insertion into Any Target Plasmid without Sequence Constraints

Exponential Megapriming PCR (EMP) Cloning-Seamless DNA Insertion into Any Target Plasmid without Sequence Constraints

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Author(s)
Ulrich, Alexander
•
Andersen, Kasper R.
•
Schwartz, Thomas
Date Issued
December 2012
Journal
PLoS ONE
Publisher
Public Library of Science
Citation
Ulrich, Alexander, Kasper R. Andersen, and Thomas U. Schwartz. “Exponential Megapriming PCR (EMP) Cloning—Seamless DNA Insertion into Any Target Plasmid Without Sequence Constraints.” Ed. Mark Isalan. PLoS ONE 7.12 (2012).
Version
Final published version
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.
MIT Department
Massachusetts Institute of Technology. Department of Biology
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Creative Commons Attribution
http://creativecommons.org/licenses/by/2.5/
Persistent DSpace Link
http://hdl.handle.net/1721.1/77196
DOI of Published Version
https://doi.org/10.1371/journal.pone.0053360
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