IOWA STATE UNIVERSITY | DEPARTMENT OF AGRONOMY | COLLEGE OF AGRICULTURE AND LIFE SCIENCES | PLANT SCIENCES INSTITUTE

1. APHIS Permit  2. Request  3. Submit  4. Plantlets or Seed

Submit your construct- Agrobacterium

  1. Transform your plasmid DNA into the Agrobacterium strain (EHA101). Please contact Kan Wang if you want to use a different strain.
  2. Extract your DNA from the Agrobacterium and check for quality using electrophoresis by including
    1. undigested DNA
    2. DNA digested with 2-3 restriction enzymes to confirm the construct.

      Restriction digestion

  3. Prepare a stab culture or plate of your DNA already in the Agrobacterium strain.
  4. Ship stab culture to PTF via overnight express (ice is not needed). The package should include:

    1. Construct map

      Stab culture or plate of bacteria, packaged as indicated in #5.
    2. Brief physical map of the construct containing
      1. major restriction enzymes positions
      2. size of the construct
    3. Gel picture illustrating the DNA quality (as described in #2
    4. Title and brief description of your project including number of events desired and type of product (seed, plant, callus).
    5. Your complete contact information including the name of PI, name of contact person for the construct communications and receiving samples, lab and office phone numbers, fax number, and e-mail addresses.
  5. Bacteria must be packaged as follows:
    1. Bacteria must be in a primary inner container (stab culture tube or bacteria plate)
    2. Secure lid with parafilm (see below)
    3. Place tube or plate in a sealable secondary container (plastic bag)
    4. Place a towel or kimwipe in the bag and seal (see below)
    5. Place bag in a cushioned envelope or box (see below)
    6. Enclose an itemized list of package contents
    7. Seal and mail

  6. You will be notified by an e-mail message of the Identification Number (ID#) that has been assigned to your construct. PTF personnel will thereafter communicate with you using this ID# rather than the original name of your construct.