MEDIUM FOR EMBRYO RESCUE (FOR 1 L)
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1808
- Put 500 mL of distilled water in a 1-L beaker.
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Add the following:
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Inorganic salts
- Muraschige�Skoog Organic medium � 34.6 g
- NaH2PO4H2O (stock 17 g/L) � 10-mL stock
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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 H2 O)
- Kinetin (for very small enbryos only) � 2 mg
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Inorganic salts
- 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.
