Scribd Logo
Upload

Welcome to Scribd!

  • 384 views

    Rye Agar (For Mycelium Growth)

    Uploaded by

    Paola Moreno López
    The document provides instructions for preparing several types of growth media for mycelium and fungi. It describes how to make rye agar by taking rye concentrate made from germinated and blended rye grains, adding sugar and agar, and autoclaving. It also provides recipes for making pea broth and agar from peas, V8-medium using V8 juice, and minimal modified Plitch medium using chemicals and glucose. The document includes concentrations of antibiotics to add to rye agar for selective media to inhibit bacterial growth.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd

    Rye Agar (For Mycelium Growth)

    Uploaded by

    Paola Moreno López
    384 views4 pages
    The document provides instructions for preparing several types of growth media for mycelium and fungi. It describes how to make rye agar by taking rye concentrate made from germinated and blended rye grains, adding sugar and agar, and autoclaving. It also provides recipes for making pea broth and agar from peas, V8-medium using V8 juice, and minimal modified Plitch medium using chemicals and glucose. The document includes concentrations of antibiotics to add to rye agar for selective media to inhibit bacterial growth.

    Original Title

    Rye agar

    Copyright

    © Attribution Non-Commercial (BY-NC)

    Available Formats

    PDF, TXT or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    The document provides instructions for preparing several types of growth media for mycelium and fungi. It describes how to make rye agar by taking rye concentrate made from germinated and blended rye grains, adding sugar and agar, and autoclaving. It also provides recipes for making pea broth and agar from peas, V8-medium using V8 juice, and minimal modified Plitch medium using chemicals and glucose. The document includes concentrations of antibiotics to add to rye agar for selective media to inhibit bacterial growth.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    384 views4 pages

    Rye Agar (For Mycelium Growth)

    Uploaded by

    Paola Moreno López
    The document provides instructions for preparing several types of growth media for mycelium and fungi. It describes how to make rye agar by taking rye concentrate made from germinated and blended rye grains, adding sugar and agar, and autoclaving. It also provides recipes for making pea broth and agar from peas, V8-medium using V8 juice, and minimal modified Plitch medium using chemicals and glucose. The document includes concentrations of antibiotics to add to rye agar for selective media to inhibit bacterial growth.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    You are on page 1of 4

    Rye agar (for Mycelium growth)

    Preparing rye concentrate 600 g rye grain (biological). Disinfect for 10 minutes in 1 litre 2% Sodium-hypochloride. Wash 4 to 5 times with distilled water using a sieve. Bring the rye over to a dry clean tub and add 500 ml distilled water. Cover the tub with Saran wrap and a dark cloth. Let the rye germinate at room temperature for 36-48 hours. Pour off the liquid en wash the grain twice with distilled water. Add 1 litre tap water and blend for 20 seconds. Pour the suspension in two 1 litre flasks Incubate in waterbath (or incubator) for 3 hours at 50C. Filter through sieve and discard grains (save suspension, about 1200 ml). Bring end volume up to 2 litre with distilled water. Store Rye concentrate at 20C

    20 % Rye Agar (RA) Take 200 ml rye concentrate. Add 20 g saccharose (table sugar) and 15 g microagar. Bring end volume with distilled water to 1 litre. Autoclave 20 minutes 120C.

    20 % Rye Agar for oospore production Take 200 ml rye concentrate. Add 20 g saccharose (table sugar), 0,1 g b-sitosterol and 15 g microagar. Bring end volume with tap water to 1 litre. Autoclave 20 minutes 120C.

    20 % Rye Agar Selective Media (RASM) Take 200 ml rye suspension. Add 20 g saccharose (table sugar) and 15 g microagar. Bring end volume with distilled water to 1 litre. Autoclave 20 minutes 120C. Let the medium cool down to luke-warm (ca. 40C). Add 10 ml antibiotic mix, and mix thoroughly. Poor the plates immediately.

    Antibiotic mix: Material: Ampicillin 250 mg/ml H2O, filter sterilised or dissolved in ethanol 70 96% Rifampicin 12 mg/ml DMSO Oxytetracycline 8 mg/ml ethanol Pimaricin 2 mg/ml methanol

    For the antibiotic mix add together: 4 ml Ampicillin, 12,5 ml Rifampicin, 6,25 ml Oxytetracyclin, 25 ml Pimaricin and 2,25 ml H2O. (Store at 20C)

    20% Rye Agar Selective Medium for fugal infections

    Take 200 ml rye suspension. Add 20 g saccharose (table sugar) and 15 g microagar. Bring end volume with distilled water to 1 litre. Autoclave 20 minutes 120C Let the medium cool down to luke-warm (ca. 40C). Add the antibiotic mix (Hohl, 1991 #3726). Antibiotics Vancomycin Polymixin B 100 mg/l 500 mg/l

    Ampicillin Rifampicin PCNB (75% WP) Benlate (50% WP) mix thoroughly.

    200 mg/l 20 mg/l 67 mg/l 100 mg/l

    Poor the plates immediately.

    Pea broth/ Pea agar (for standard mycelium growth)


    Autoclave (15 minutes 120C) 120 g frozen peas (supermarket) in 1 litre distilled water. Strain peas from pea broth using 4 layers of cheese clothe squeezing gently to remove all excess liquid from peas. Bring volume of broth to 1 litre. For pea agar add 15 g agar to 1 litre broth. Autoclave for 20 minutes 120C.

    V8-medium (for mycelium growth)


    Combine 200 ml V8 juice and 800 ml tapwater. Add 2 g CaCO3 and 0.05 g b-sitosterol and mix well. Add 15 g agar. Autoclave for 20 minutes at 120 C. Stir medium well before poring the plates in order to resuspent the CaCO3.

    Modified Plitch medium (minimal medium for mycelium growth)

    Prepare for 1 litre medium: 0.5 g KH2PO4 0.25 g Mg2SO4.7H2O 1.0 g L- aspergine 0.5 g Yeast extract 0.01 g b-sitosterol 25.0 g Glucose 0.001 g Thiamine 15 g microagar

    Bring volume to 1 litre with Milli Q water Autoclave for 20 minutes at 120 C.

    You might also like