We are using HgCl2 at two stages of surface sterilization of banana shoots as 1mg/l for 10 minutes outside the LAF and finally 1mg/l for 5 minutes inside LAF. We are getting excess tissue blackening and also considering the rate of HgCl2 (INR:3200/100g), please suggest me any of the effective substitute for the same.
Domestic bleach ACE, diluted 1:1. Time of treatment needs to be adjusted
Original banana shoot cuttings (from the mother tree) are sterilized by immersion on sodium hypochlorite (5%) for 20 min followe by 5 min in mercuric chloride (0.1 %), the rinse by sterilized H2O for 3 successive times (5, 10 and 15 min, respectively).
To reduce the blacking in tissue culture medium, there are three ways and you can use one of them. the 3 ways are: 1) you may place the explants on multiplication medium supplemented with 1g/L charcool; 2) add an antioxidant the the medium, antioxidants such as ascorbic acid (200 mg/L), PVP (200 mg/L), or citric acid; 3) you should subculture the explants many times at the begining of culture.
you can use sodium hypochoride , calcium hypochloride, hydrogen peroxide.
It’s a good technical Question
You can use sodium hypochlorite (2.5%) for 5 min followed by hydrogen peroxide (1.0 %), for 5 min or till no bubbles are observed finally the rinse by sterilized distilled water 3 successive times (5, 10, and 15 min, respectively).
good luck
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Methods
- Cloning