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Wheat Genetics Resource Center

Wheat Genetics Resource Center
Kansas State University

Throckmorton Plant Sciences Center
Department of Plant Pathology
Manhattan, KS 66506-5502
785-532-2366 (TEL)
785-532-5692 (FAX)
wgrc@k-state.edu

Kansas Wheat Innovation Center

1990 Kimball Ave
Manhattan, KS 66502
785-320-4383 (TEL)

 

MEDIUM FOR EMBRYO RESCUE (FOR 1 L)

  1. Put 500 mL of distilled water in a 1-L beaker.
  2. Add the following: 
    • Inorganic salts
      • Muraschige–Skoog Organic medium – 34.6 g
      • NaH2PO4H2O (stock 17 g/L) – 10-mL stock
    • Organic constituents 
      • ucrose (6 % final concentration) – 30 g
      • i-inositol (100 mg/L stock is 0.5 g/50 mL) – 10 mL
      • Thiamine (0.4 g/l stock is 2 g/50 mL) – 10 mL
      • Amino acids L-tyrosine, L-argenine, glycine – 10 mL (10 mg/L stock is 50 mg each aa/50 mL H2O)
      • Kinetin (for very small enbryos only) – 2 mg
  3. Dilute to 800 mL.
  4. Adjust pH to 5.7 with NaOH..
  5. Bring volume to 1,000 mL.
  6. Add 8 grams Bacto® agar.
  7. Melt agar in microwave for 3 minutes or dissolve while stirring on a hotplate.
  8. Pour ~20 mL into Kimax culture tubes (50 mL) and cap.
  9. Sterilize by autoclaving for 15 minutes.
  10. Slant in tubes or pour into petri dishes.