I have been trying to grow Nicotiana benthamiana via tissue culture. I used ethanol and sodium hypochlorite with different concentrations and time duration for seeds sterilization and lastly washed the seeds with distilled water but there is no growth.
At some concentrations there is no contamination but the seeds didn't respond.
Currently, I have only Ethanol and sodium hypochlorite as sterilizing agents.
Could anyone help me to grow N.benthamiana using these agents?
Thanks
Regards
Nadeem Ahmad
I use sodium hypochlorite 0,1% for 15 min and lastly washed the seeds with distilled water (5/6 time )
Try adding in a bit of surfactant with the bleach/ethanol step. If you have tween or silwet those work well, even dish soap can help. It doesn't take much, try 0.05% as a starting point.
Do your seeds germinate well in the absence of surface sterilization? If not, it might be time for a fresh batch of seeds.
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