Some tissues are more difficult than others for establishing clean cultures. The concentration of chlorine bleach and the length of time the tissue is in the bleach can vary. Delicate, succulent tissue may be clean after 10 min in a 10% solution. Fern spores may only require a 3-5% bleach for 3-4% min. Some seeds may require 30-50% bleach for 20-60 min. Any cut explant such as a stem or leaf that is surface sterilization. These damaged pieces should be removed before culture.
Contamination that results from improperly sterilized tissue will generally arise from the explant and be located in the medium adjacent to explant. Contamination due to poor technique generally will appear over the entire agar surface. Contamination of cultures by fungi appears as a fuzzy growth. Bacterial contamination appears as smooth pink, white, or yellow colonies. If contamination is due to poor technique, contamination transfer hood filters, or improperly sterilized media it will be scattered on or in the medium. Contamination from insects will generally appear as tracks across the medium, which are visible due to bacterial or fungal growth on the insect tracks. For a more detailed discussion on contamination in vitro, refer to Chaper5
Some tissues are more difficult than others for establishing clean cultures. The concentration of chlorine bleach and the length of time the tissue is in the bleach can vary. Delicate, succulent tissue may be clean after 10 min in a 10% solution. Fern spores may only require a 3-5% bleach for 3-4% min. Some seeds may require 30-50% bleach for 20-60 min. Any cut explant such as a stem or leaf that is surface sterilization. These damaged pieces should be removed before culture. Contamination that results from improperly sterilized tissue will generally arise from the explant and be located in the medium adjacent to explant. Contamination due to poor technique generally will appear over the entire agar surface. Contamination of cultures by fungi appears as a fuzzy growth. Bacterial contamination appears as smooth pink, white, or yellow colonies. If contamination is due to poor technique, contamination transfer hood filters, or improperly sterilized media it will be scattered on or in the medium. Contamination from insects will generally appear as tracks across the medium, which are visible due to bacterial or fungal growth on the insect tracks. For a more detailed discussion on contamination in vitro, refer to Chaper5
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