Transient Transformation of Plant Protoplasts
Corresponding Organization :
Other organizations : Vlaams Instituut voor Biotechnologie, Ghent University
Variable analysis
- Amount of pCDB-Cas9-GFP-PDS (10 or 20 μg)
- Amount of pKAR6 (10 or 20 μg)
- Transfection efficiency (ratio of GFP-expressing protoplasts to total living protoplasts)
- Protoplast culture conditions (dark, 23 ± 2°C, 30 rpm, 20 h)
- Centrifugation parameters (5 min at 80 g)
- Microscopy settings (Zeiss AxioImager M2, Axiocam MRm camera, ZEN software, 200× magnification)
- Positive control: Protoplasts transfected with pKAR6 (expressing GFP under 35S promoter)
- Negative control 1: Protoplasts transfected without vector
- Negative control 2: Protoplasts without the addition of both PEG and vector
Annotations
Based on most similar protocols
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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