Jan 06, 2025
Transformation of chemically competent Agrobacterium cells (freeze-thaw method)
This protocol is a draft, published without a DOI.
- 1CSJMU, Kanpur
Protocol Citation: Velu Mani Selvaraj 2025. Transformation of chemically competent Agrobacterium cells (freeze-thaw method). protocols.io https://protocols.io/view/transformation-of-chemically-competent-agrobacteri-dw6v7he6
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: In development
We are still developing and optimizing this protocol
Created: January 06, 2025
Last Modified: January 06, 2025
Protocol Integer ID: 117685
Abstract
Protocol to transform chemically competent Agrobacterium cells
Materials
YEM plates
Water bath
Liquid nitrogen
Add about ~1 µg of plasmid DNA into the 100 µl of competent cell. Freeze the cells by placing in liquid nitrogen.
Immediately thaw cells by transferring to 37 ℃ for 5 min.
Add 1 ml of YEM to the cells and incubate at 28 ℃ for 4 hrs.
Spread the cells (upto 200 µl/plate) on YEM agar with appropriate antibiotic.
Incubate the cells at 28 ℃ for 1 - 3 days to check for transformants.