I have a new problem now. I tried 3 protocols but each time contamination occurred.
Cleaning with tap water for 30 minutes before each protocol.
1. 70% Etoh 5 min + 1.5% domestos 10 min negative result + dead brown leaves.
2. 70% Etoh 4 min + 4% domestos 8 min + 3.5% domestos 5 min negative result + dead brown leaves.
3. 10% NaCIO 15 min negative result + leaves are alive.
Hi,
which protocol are you already using? It would be better to know what protocol are you following to help you to further understand the troubles.
Here are some useful links:
Article An efficient plant regeneration protocol from callus culture...
https://www.twjeas.org/PDFs/VOL%203%20ISSUE%202%20(MARCH)%202017/Regeneration%20of%20plantlets%20from%20in%20vitro%20cultured%20seed%20of%20endangered%20Aegle%20marmelos%20(L.)%20Corr.%20Rutaceae.pdf
https://www.hindawi.com/journals/aag/2020/8840386/
Article In Vitro shoot organogenesis and regeneration of plantlets f...
Best regards
Uğur Kaya let us know your protocol, if you are using the sodium hypochlorite use HgCl2 0.1% solution otherwise reduce the sodium hypochlorite working concentration and increase the number of wash.
@Zahid Fazal , @francesco Desiderio and @Ram Krishna thank you for answering me.
I have a new problem now. I tried 3 protocols but each time contamination occurred.
Cleaning with tap water for 30 minutes before each protocol.
1. 70% Etoh 5 min + 1.5% domestos 10 min negative result + dead brown leaves.
2. 70% Etoh 4 min + 4% domestos 8 min + 3.5% domestos 5 min negative result + dead brown leaves.
3. 10% NaCIO 15 min negative result + leaves are alive.
I noticed you wrote that you are using tap water for your protocol. Do you mean DI water that has been autoclaved? Or is it straight from the sink? If you are not using DI water that can be the cause of the contamination.
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