SLIC: a method for sequence- and ligation-independent cloning

Methods in Molecular Biology
Mamie Z Li, Stephen J Elledge

Abstract

We describe here a method for sequence- and ligation-independent cloning (SLIC). SLIC uses an exonuclease, T4 DNA polymerase, to generate single-stranded DNA overhangs in insert and vector sequences. These fragments are then assembled in vitro and transformed into Escherichia coli to generate recombinant DNA of interest. SLIC inserts can also be generated by incomplete PCR (iPCR) or mixed PCR. As many as five inserts can be assembled in one reaction simultaneously with great efficiency using SLIC. SLIC circumvents sequence constraints for recombinant DNA using standard restriction enzyme-mediated cloning and previous ligation-independent cloning methods and provides a new approach for the efficient generation of recombinant DNA.

Citations

Oct 21, 2015·Proceedings of the National Academy of Sciences of the United States of America·Amy B BantaPaula V Welander
Oct 22, 2016·ACS Synthetic Biology·Simon John MoorePaul S Freemont
Jul 13, 2016·Proceedings of the National Academy of Sciences of the United States of America·Mary E Wahl, Andrew W Murray
Oct 18, 2016·ACS Synthetic Biology·Ryan M PhelanJay D Keasling
Dec 29, 2016·Proceedings of the National Academy of Sciences of the United States of America·Amy B BantaPaula V Welander
Jan 25, 2017·Proceedings of the National Academy of Sciences of the United States of America·Cheng Wang, Stephen P Goff
Feb 9, 2017·Genes & Development·Sida LiaoStephen J Elledge
Apr 13, 2017·The Journal of Cell Biology·Gantas Perez-MockusFrançois Schweisguth
Mar 28, 2017·Nature Chemical Biology·Christian RothGideon J Davies
Mar 10, 2017·Human Mutation·Philippe LemayZoha Kibar
Mar 21, 2017·Nature Chemical Biology·Marta C MarquesInês A C Pereira
Mar 2, 2017·Cold Spring Harbor Perspectives in Biology·Christian R BoehmJim Haseloff
Sep 8, 2017·Biomolecular NMR Assignments·Edith BuchingerFinn L Aachmann
May 15, 2018·Antioxidants·Evangelos ZoidisGeorgios P Danezis
Apr 11, 2018·The Journal of Biological Chemistry·Matthew Blain-HartungJ Clark Lagarias
Feb 20, 2018·Molecular Neurobiology·Eun-Hye HongJeong-Yoon Kim
Apr 2, 2014·Infection and Immunity·Antonino MissineoGiampiero Pietrocola
Mar 1, 2015·Applied and Environmental Microbiology·Steven D SmithJudy D Wall
Apr 14, 2016·PloS One·Roger M BenoitManfred Auer
Jul 16, 2015·PloS One·Christina S NødvigUffe H Mortensen
Feb 8, 2019·Biotechnology and Bioengineering·Liping WangYunzi Luo
Aug 19, 2015·Nature Chemical Biology·Eunji KimYeo Joon Yoon
Oct 16, 2013·Journal of Industrial Microbiology & Biotechnology·Ryan E CobbHuimin Zhao
Jan 4, 2015·The EMBO Journal·Sara K PowellDavid M MacAlpine
Apr 5, 2014·Acta Crystallographica. Section F, Structural Biology Communications·Juha Pekka Kallio, Inari Kursula

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