What is the List of Requirement of AME'S Test

                                                           AME’s Test Requirement:

Chemicals:

        1            Distilled water

        2            Dextrose

        3            Magnesium sulfate (MgSO₄.H₂O)

        4            Citric acid monohydrate

        5            Potassium phosphate dibasic anhydrous (K₂HPO₄)

        6            Sodium ammonium phosphate (Na₂NH₂PO₄.4H₂O)

        7            D-biotin (F.W. 247)

        8            L-Histidine (F.W. 191.7)

        9            Sodium chloride

    10            Oxoid nutrient broth

    11            Sodium phosphate monobasic

    12            Sodium phosphate dibasic

    13            D-glucose-6-phosphate

    14            Nicotinamide adenine dinucleotide

    15            Phosphate (NADP)

    16            Magnesium chloride (MgCl)

    17            Potassium chloride (KCl)

    18            Glucose-5-phosphate dehydrogenase

    19            Liver S-9 fraction

    20            Flavin mononucleotide (FMN)

    21            Ampicillin

    22            Tetracycline

    23            Crystal violet

Glassware’s/General requirement

 

        1            Petri plate (100×15-mm)

        2            Sterile glass tubes (100×13-mm and 150×16-mm)

        3            Test tube racks

        4            General laboratory glassware (flasks, bottles, graduated cylinders)

        5            Pipets (1, 2, 5, and 10 ml)

        6            Pipettors (adjustable volumes up to 200 and 500 ml)

        7            Pipet tips

        8            Dispensers for delivering top agar, buffer, and S-9 mix to the test tubes

        9            Cryogenic tubes for freezing down permanent and working cultures

    10            Sterile glycerol

    11            Magnetic stir bars

    12            Mammalian tissue homogenate (S-9 fraction)

    13            Dimethylsulfoxide and other solvents

    14            Positive control chemicals

    15            Gloves

    16            Gowns

    17            Goggles or protective eyewear Biohazard bags

    18            Protective covers for workspace

 

Equipment

        1            Colony counter (manual or electronic)

        2            Autoclave

        3            Shaking incubator set at 120 rpm and 37°C for growing the overnight cultures

        4            Incubator for incubating the GM agar plates equipped with electrical outlet for Desiccator assays

        5            Oven, heating or water bath set at 43°C to 48°C to maintain the temperature of top agar

        6            Gas line for keeping aseptic techniques while inoculating cultures

        7            Boiling water bath or microwave oven for melting top agar.

        8            Desiccator (9 liter)

        9            Magnetic stirrers

    10            Balances

    11            Water purification system to generate distilled water

    12            Spectrophotometer

    13            Centrifuge (up to 8000 rpm)

    14            Revco freezer or liquid nitrogen tank

    15            Refrigerator/freezer

    16            Chemical/biological safety hood

    17            Liquid waste disposal

    18            Vacuum pump

    19            Solid waste disposal

 

Reagents and media

        1            Vogel Bonner (VB salt medium ) E 50x

        2            Glucose solution (10% v/)

        3            GM agar plates

        4            Histidine/biotin solution (0.5 mM)

        5            Sodium phosphate buffer 0.1 mM, ph 7.4

        6            Cofactors for S-9 mix

        7            Ampicillin solution (0.8 % w/v)

        8            Tetracycline solution (0.1 % w/v)

Bacterial cultures

 

        1            Salmonella typhimurium TA98

        2            Salmonella typhimurium TA100

        3            Salmonella typhimurium TA1535

        4            Salmonella typhimurium TA1537 or TA97 or TA97a

        5            Salmonella typhimurium TA102 or Escherichia coli WP2 uvrA or Escherichia coli  WP2 uvrA (pKM101)

        6            Post mitochondrial supernatant of rat liver homogenate (S-9)

 

Controls

 

        1            9-Aminoanthracine

        2            4-Nitro-o-phenylenediamine

        3            Sodium azide

        4            Mitomycin C

        5            Methyl methane sulfonate

        6            9-aminoacridine

        7            2-Aminoanthracine

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