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Updated: Nov 23, 2025
Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes
Published on: February 14, 2020
BiBAC Modification and Stable Transfer into Maize (Zea mays) Hi-II Immature Embryos via Agrobacterium-Mediated
Jon P Cody1, Nathaniel D Graham1, James A Birchler1
1Division of Biological Sciences, Tucker Hall, University of Missouri, Columbia, Missouri.
Abstract:
Binary Bacterial Artificial Chromosomes (BiBAC) are large insert cloning vectors that contain the necessary features required for Agrobacterium-mediated transformation. However, the large size of BiBACs and low-copy number in Escherichia coli (DH10B) and Agrobacterium tumefaciens make cloning experiments more difficult than other available binary vector systems. Therefore, a protocol that outlines preparation, modification, and transformation of high-molecular weight (HMW) constructs is advantageous for researchers looking to use BiBACs in plant genomics research. This unit does not cover the cloning of HMW DNA into BiBAC vectors. Researchers looking to clone HMW DNA into BiBACs can refer to Zhang et al. (2012; doi: 10.1038/nprot.2011.456). © 2017 by John Wiley & Sons, Inc.
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