发布时间:2019-08-16 17:18 原文链接: MEDIUMFOREMBRYORESCUE(FOR1L)

  1. Put 500 mL of distilled water in a 1-L beaker.

  2. Add the following:

    • 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

    • Muraschige–Skoog Organic medium – 34.6 g

    • NaH2PO4H2O (stock 17 g/L) – 10-mL stock

    • Inorganic salts

    • Organic constituents

    • Dilute to 800 mL.

    • Adjust pH to 5.7 with NaOH..

    • Bring volume to 1,000 mL.

    • Add 8 grams Bacto® agar.

    • Melt agar in microwave for 3 minutes or dissolve while stirring on a hotplate.

    • Pour ~20 mL into Kimax culture tubes (50 mL) and cap.

    • Sterilize by autoclaving for 15 minutes.

    • Slant in tubes or pour into petri dishes.