Adding Phenol Red to plant tissue culture media?

[MC]

Hello, after doing some reading on the effects of autoclaving and callus to TC agar media pH, I've come to the conclusion that the actual pH can vary quite wildly with time compared to the measured pH prior to autoclaving. I was thinking if an indicator like phenol red can be added to the media to act as a constant pH indicator. Would this idea work?

[Sebastian S Cocioba]

It gets taken up and does weird things to seedlings. Just use MES buffer at 0.5g/L to handle the variance over time since sucrose metabolism acidifies the media. No need for indicators though it us fun to see the uptake. I played with universal buffers and chlorophenol red. 

Sebastian S. Cocioba

CEO & Founder

New York Botanics, LLC

Blog: ATinyGreenCell.com

On Jan 9, 2017, at 3:06 PM, MC <marky...@gmail.com> wrote:

Hello, after doing some reading on the effects of autoclaving and callus to TC agar media pH, I've come to the conclusion that the actual pH can vary quite wildly with time compared to the measured pH prior to autoclaving. I was thinking if an indicator like phenol red can be added to the media to act as a constant pH indicator. Would this idea work?

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[Bryan Jones]

What are you growing? Phenol red is often included in mammalian tissue culture media for exactly that purpose. But it does have some effects on the cells since it's a weak estrogen analog (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC323325/).

To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/01AD2CB5-8BD8-46D3-8376-AB1F8056F5A2%40gmail.com.

[MC]

Thanks for the responses.

What is everyone's experience with TC and pH? What protocol do you follow for pH adjustment or buffering? I've read the literature on pH, but I'm curious if it really means life or death for the culture as some sources seem to portray.