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  Section: Biotechnology Methods » Tissue Culture Techniques
 
 
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Media for Plant Tissue Culture

 
     
 
Content
Tissue Culture Techniques
  Tissue Culture Methods
  Plant Tissue Culture
  Plant Tissue Culture (Cont.)
  Many Dimensions of Plant Tissue Culture Research
  What is Plant Tissue Culture?
  Uses of Plant Tissue Culture
  Plant Tissue Culture demonstration by Using Somaclonal Variation to Select for Disease Resistance
  Demonstration of Tissue Culture for Teaching
  Preparation of Plant Tissue Culture Media
  Plant Tissue Culture Media
  Preparation of Protoplasts
  Protoplast Isolation, Culture, and Fusion
  Agrobacterium Culture and Agrobacterium — Mediated transformation
  Isolation of Chloroplasts from Spinach Leaves
  Preparation of Plant DNA using
  Suspension Culture and Production of Secondary Metabolites
  Protocols for Plant Tissue Culture
  Sterile Methods in Plant Tissue Culture
  Media for Plant Tissue Culture
  Safety in Plant Tissue Culture
  Preparation of Media for Animal Cell Culture
  Aseptic Technique
  Culture and Maintenance of Cell Lines
  Trypsinizing and Subculturing Cells from a Monolayer
  Cellular Biology Techniques
  In Vitro Methods
  Human Cell Culture Methods

This will help prevent senescence, which can occur when cultures are grown on 1 medium only.


Fertilizer Stock Mix
  • ½ tablespoon all-purpose 18:18:18 (N.P.K.) water-soluble fertilizer
  • 2 cups of water
  • Gently boil to dissolve all ingredients and store in the refrigerator
Figure 13
Basic Nutrient Mix
  • 2 cups of distilled water
  • Between 2 tablespoonfuls and ¼ cup of sugar
  • ½ tablespoon of fertilizer stock
  • ½ Inositol tablet (500 mg)
  • ½ vitamin tablet with thiamine
  • Gently boil to dissolve all ingredients and store in the refrigerator
Rooting Mix
  • ½ teaspoon of malt
  • Add basic nutrient mix to make 2 cups
  • Check the pH using narrow-range pH indicator tape
  • Adjust pH to between 5 and 6 with vinegar or sodium bicarbonate
  • Add several drops of yellow food coloring to identify the formula
  • Gently boil to dissolve all ingredients and cool
  • Cover and put in the glove box
  • Add 100 mg silver nitrate
  • Pour into sterile jars and cap
  • Store in the refrigerator until use
Multiplication Mix
  • ¼ to ½ cup of coconut milk
  • Add basic nutrient mix to make 2 cups
  • Check the pH using narrow-range pH indicator tape
  • Adjust pH to between 5 and 6 with vinegar or sodium bicarbonate
  • Add several drops of red food coloring to identify the formula
  • Gently boil to dissolve all ingredients and cool
  • Cover and put in the glove box
  • Add 100 mg silver nitrate
  • Pour into sterile jars and cap
  • Store in the refrigerator until use
Peroxidated Flowering Mix
  • (Optional) ½ teaspoon of malt
  • Add basic nutrient mix to make 2 cups
  • Add several drops of blue food coloring to identify the formula
  • Sterilize and cool to around 55°C
  • Cover and put in the glove box
  • Stir in ½ teaspoon peroxide
  • Add 100 mg silver nitrate
  • Pour into sterile jars and cap
  • Store in the refrigerator until use
Sand-Supported Culture Protocol
The purpose of agar in growth media is solely to support the tissue. Instead of agar, use sand to support the culture.
  1. Boil sand in water on your stovetop.
  2. Store the clean sand in a plastic bag.
  3. Put 1 ounce of sand (volume) in a baby food jar.
  4. Sterilize jars in your pressure cooker.
  5. Place sterilized jars, prepared caps, and sterile media solution in your glove box.
  6. In the glove box, add enough sterile media solution to just saturate but not cover the sand.
  7. Close the jar with a prepared cap.
  8. Store in the refrigerator until use.
    Note:
    Do not microwave the sand, it will splatter all over.
    The following are bacterial or fungal agars, use them for ideas.
Amaranth Soy Agar
  • 40 grams amaranth flour
  • 40 grams soy flour
  • 19 grams agar
  • 1 liter distilled water
Entheo Genesis No. 442
  • 20 grams amaranth flour
  • 20 gm brown rice flour
  • 20 gm potato flour
  • 20 gm soy flour
  • 4 gm malted barley
  • 19 gm agar
  • 1 liter distilled water
Cornmeal Dextrose Agar
  • 50 gm yellow cornmeal
  • 5 gm dextrose
  • 19 gm agar
  • 1 liter distilled water
Barley Malt Extract Agar
  • 40 gm barley flour
  • 2 gm malt extract
  • 1–2 gm yeast extract (optional)
  • 9.5 gm agar
  • 500 liter distilled water
Dr. Pollock’s Modified Agar
  • 20 gm dried dog food
  • 20 gm amaranth flour
  • 4 gm dextrose or malt extract
  • 1 liter distilled water
Malt Extract Peptone Agar
  • 30 gm malt extract
  • 2 gm soya peptone
  • 15 gm agar
  • 1 liter distilled water
  • Adjust pH to 5.6. Sterilize for 16 min.
Malt Agar
  • 20 gm of agar
  • 20 gm of malt extract
  • 2 gm of yeast
  • Water to 1 liter
Potato Dextrose Agar
  • 20 gm of agar
  • The strained broth produced by boiling 300 grams of diced potatoes in 1 liter of water for 1 hour
  • 10 grams of dextrose
  • Clean water back up to 1-L volume
  • In one experiment, tapioca starch produced the greatest number of shoots and nodes.

 
     
 
 
     




     
 
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