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        Yeast Plates

        互联网

        1035

         

        Yeast Plates

        Hahn Lab

        Last modified Tue, Oct 6, 1998

        Amino Acid Mix (40X ) Use 0.6 g/liter
        Tyrosine 2 g
        Arginine 4 g
        Serine 2 g
        Valine 2 g
        Threonine 4 g
        Isoleucine 2 g
        Phenylalanine 2 g
        Aspartic Acid 2 g
        Proline 2 g

         


        Glucose Complete Plates (1 liter):

        1.7 g Yeast Nitrogen base -amino acids and ammonium sulfate
        5.0 g Ammonium Sulfate
        1 liter H2O
        pH to 7.0 using NaOH

        Add:
        0.6 g Amino Acid mix (mix well before adding)
        20g Bacto-agar (Difco)

        Autoclave 25 min.

        Add:
        10ml (10/mg/ml) of any other required amino acids
        10ml 0.4% Adenine sulfate in 0.1M HCl if required
        10 ml 0.2% Uracil if required
        50 ml 40% glucose

         


        YPD Plates (1 liter):

        Yeast Extract 10 g
        Bacto peptone 20g
        Bacto-Agar 20g
        H2O 1 liter

        Autoclave 25 min

        Add:
        50 ml 40% glucose
        5 ml 0.4% Adenine sulfate in 0.1 M HCl if required
        Add 10 ml 0.2% Uracil (only if plates will be used for tetrad dissection)

         


        5-FOA plates (0.5 liter):

        Agar:

        15 g Bacto-agar
        300 ml H20 in 500 ml flask
        Autoclave 25 min.
         

        FOA mix:

        0.85 g Yeast Nitrogen base - amm. sulfate and amino acids
        2.5 g Ammonium sulfate
        0.3 g amino acid mix (from above)
        25 ml 40% glucose
        17.5 mg Uracil
        0.5 g 5-FOA
        Any other required amino acids
        225 ml H2O

        Microwave the above mixture for about 3 min until about 65 degrees. Stir on stir plate until dissolved (~10-15 min). Filter through 0.2 micron bottle top filter into sterile bottle. Add to autoclaved agar. Mix well and pour plates. Do not pH the plates as this will inactivate the 5-FOA.

         

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