Protoplast Transient Assay in Arabidposis

Question

[Maryam]

I am currently working on establishing a protocol for salt stress treatments in Arabidopsis protoplasts for transient expression assays,I would greatly appreciate your expertise on some specific aspects of the protocol.

Regarding salt stress treatments:

• What NaCl concentration do you recommend for optimal stress response in Arabidopsis protoplasts without causing excessive cell death?
• What is the optimal incubation time for salt stress treatments in your experience?

I have to incubated for 16 hr and then Add 650ul of W5 to “control” wells and 650ul of W5 (0.32M NaCl) to “stress” wells .

For protoplast transformation:

What is the optimal DNA concentration (plasmid) for PEG-mediated transformation of protoplasts in salt stress experiments?

[dthorbur]

https://biology.stackexchange.com/ might be a better place to go for wet lab questions.

[colindaven]

This is a bioinformatics site so you are unlikely to get a good answer here - it is way off topic :-)