I already try to sterile my fern for in vitro spore culture by:-
1. wash under running tap water
2. then shake with fungicide + distill water
3. rinse with distill water(3 times)
4. dip in 1 min 70% ethanol
5. dip in difference concentration of sodium hypochlorite (if higher concentration the it cannot grow but lower concentration cause fungus)
6. rinse with distill water (3 times)
You can read : An Efficient Method for Surface Sterilization and Sowing Fern Spores in vitro
Article An Efficient Method for Surface Sterilization and Sowing Fer...
In vitro spore germination and gametophyte growth assessment of a critically endangered fern: Pteris tripartita Sw
Article In vitro spore germination and gametophyte growth assessment...
Hello Nurul,
thanky for posting this very interesting technical question on RG. In my opinion you did everything right. The following link has a detailed protocol for sterilization of fern spores using sodium hypochlorite solution:
Surface Sterilization of C-Fern Spores
http://www1.biologie.uni-hamburg.de/b-online/library/cfern/cfern.bio.utk.edu/manual/cfmsterilespore.html
Mercuric chloride (HgCl2) is also frequently used for this purpose. However, I would strongly suggest to avoid working with this compound if at all possible because of the very high toxicity of mercury and its compounds.
Good luck with your research!
0.1% mercuric chloride solution prepared in autoclaved distilled water can be used for surface sterilization of fern spores.After soaking for about 5min,the explants are washed number of times thoroughly with sterile distilled water before inoculation.
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