I am not observing uniform induction of my promoter fused to GUS in xylem vasculature of taproot of tomato.
Key questions are whether I should do cross-sections after or before putting in X-gluc solution? If I put whole taproot in X-gluc solution will it absorb enough for reaction, considering the presence of exodermis? Also, for how much time do I need to keep in Acetone?
Thanks in advance.
Anurag
It could include other evaluations, for example evaluating the treatment (s) on physicochemical parameters or trace elements in stems and roots. With ICP-MS analysis and FTIR analysis you could know changes in the elements in the plants. These parameters could determine the effect of the treatments.
Hi Anurag,
GUS staining is spatiotemporal. Its intensity varies with the construct and the promoter activity in the tissue-specific level (you need to standardize the conditions for your transgenic lines).
First, perform the GUS staining with a vacuum (I use 20 mins for my samples, you can try different conditions), followed by several 70% ethanol washes, and then do the cross-sectioning.
I use 70% ethanol to wash several times because I use leaf tissue (to wash off chlorophyll).
Good luck with your work.
Thank you very much for your valuable answers. All of them have been very helpful for me J. C. Tarafdar Hermes Pérez-Hernández Dharmesh Jain
- Badges
- Science topic
- Similar topics
- Agricultural Science
- Agronomy
- Crop Science
- Crop