I'm planning to do a stable transformation of BY-2 cells using Agrobacterium tumefaciens C58C1 with plasmid containing hygromycin resistance gene marker. What concentration of hygromycin I should use to select transgenic BY-2 calli?
Hello, you can use 30ug/mL hygromycin B on 0.8% agar plates.
Good luck!
You should contact the authors from the following paper (2018) to find out. They were doing the similar things as you do--using BY-2 cells for transformation, then use hygromycin (hpt gene in their binary vectors) for selection (see attachment, Figure 2. construct description).
Article Establishment of a tobacco BY2 cell line devoid of plant spe...
Hello, you can use 30ug/mL hygromycin B on 0.8% agar plates.
Good luck!
@ Karen A. Darbinyan, is the 30 ug/mL concentration reported for BY-2 calli? If not, calli derived from different tobacco varieities (ex. W38, Little Harvana,...) can have different degrees of tolerance towards hygromycin B.
Karen A. Darbinyan Thank you for the concentration.
Yuan-Yeu Yau I'm planning to contact the authors.
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