List of Laboratory Requirements for AME's Test

 

AME’s Test

Requirement:

Chemicals:

       1          Distilled water

       2          Dextrose

       3          Magnesium sulphate (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 plates (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 eye wear Biohazard bags

    18          Protective covers for work space

 

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 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-phenylenediame

       3          Sodium azide

       4          Mitomycin C

       5          Methyl methane sulfonate

       6          9-aminoacridine

       7          2-Aminoanthracine

                                                            END OF THE DOCUMENT

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