MegaGate: A toxin-less gateway molecular cloning tool.

Journal Article (Journal Article)

Gateway cloning employs the use of the ccdb toxin and has low colony numbers, making it difficult to apply at scale to clone libraries of cDNA vectors. In this protocol, we describe MegaGate, a toxin-less Gateway technology capable of robust cDNA library cloning that is efficient, cheap, and scalable. MegaGate eliminates the ccdb toxin used in Gateway recombinase cloning and instead utilizes meganuclease-mediated digestion to eliminate background vectors during cloning and is 99.8% efficient with high colony numbers. For complete details on the use and execution of this protocol, please refer to Kramme et al. (2021).

Full Text

Duke Authors

Cited Authors

  • Kramme, C; Plesa, AM; Wang, HH; Wolf, B; Smela, MP; Guo, X; Kohman, RE; Chatterjee, P; Church, GM

Published Date

  • December 2021

Published In

Volume / Issue

  • 2 / 4

Start / End Page

  • 100907 -

PubMed ID

  • 34746865

Pubmed Central ID

  • PMC8551244

Electronic International Standard Serial Number (EISSN)

  • 2666-1667

International Standard Serial Number (ISSN)

  • 2666-1667

Digital Object Identifier (DOI)

  • 10.1016/j.xpro.2021.100907


  • eng