Plant Transformation
Location
Morgenstelle 32
5. Stock, Raum 5X17
phone: 73223
Booking email: transformation@zmbp.uni-tuebingen.de
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Equipment
2 sterile benches, 3 growth rooms
| 1. The unit offers Agrobacterium-mediated tissue-culture transformation of potato, tobacco and tomato ssp. |  |
| 2. PEG-mediated transformation of protoplasts from cell cultures or root cells (Arabidopsis). |  |
| 3. Maintenance of our Col-0 cell culture (wild type). |  |
Examination of the protoplat transformation efficiency by flow cytometric anaylsis (FCA).
Protoplasts isolated from root cell culture are transformed with a 35S::GFP construct (12kb) as a routine control. As an alternative to calculating the transformation efficiency, we have used FCA as a means for detecting cells expressing GFP. The negative population is marked in black. Cells that are strongly expressing GFP have a fluoresence signal that is well above that of the total plant cell population‘s green autofluoresence (cells marked in dark green); cells marked in light green are those with weaker GFP fluoresence emission. In this example 23% of the cells were transformed with GFP, and 50% of those are strongly expressing GFP.