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
Language
- eng