How to Make a Brassica Tissue Culture

Tissue culture is a quick method for asexually propagating large numbers of plants in a relatively short period of time. The resulting plants will be clones of the original source. The process must be sterile from the outset, and the procedure must take place within a transfer hood in a tissue culture lab. Various lab containers may be used to hold the agar plus nutrient substrate that will provide the initial growth environment for the transplants, such as petri dishes.

Things You'll Need

  • Sterile tissue culture medium in sterile petri dish
  • High-purity water
  • Sterile forceps
  • Sterile scalpel
  • Bunsen burner
  • Flint striker
  • Small beaker with 70 percent ethanol (EtOH)
  • Squeeze bottle with 70 percent EtOH
  • Sterile paper towels
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Instructions

    • 1

      Use brassica plants grown from aseptic seed germination (seeds that have undergone a sterilization procedure prior to germination), or plants that have undergone specific sterilization protocols and been tested for lack of microbial contamination.

    • 2

      Wash your hands and wipe them with 70 percent ethanol, then apply gloves. Wipe fume hood surfaces with 70 percent ethanol. Place the petri dish containing agar, supplemented with nutrients, into the fume hood after wiping the upper and lower surface of the dish with 70 percent ethanol.

    • 3

      Place the plant between two sheets of sterile paper towels. Cut a five-to-10 millimeter piece of plant from any part of the plant with the scalpel. Keep the scalpel and forceps in the beaker of ethanol when not in use.

    • 4

      Use the forceps to place the cut piece of plant into the sterile petri dish and press the cut end into the nutrient-rich agar.

    • 5

      Incubate the transplants in a growth chamber or greenhouse until rooted.

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