What is the List of Requirement of In-vitro Mammalian Chromosome Aberration Test?

 In-vitro Mammalian Chromosome Aberration Test

Requirement:

Chemicals:

        1            Distilled water

        2            Magnesium sulfate (MgSO₄.H₂O)

        3            Potassium phosphate dibasic anhydrous (K₂HPO₄)

        4            Sodium ammonium phosphate (Na₂NH₂PO₄.4H₂O)

        5            Sodium chloride

        6            Sodium phosphate monobasic

        7            Sodium phosphate dibasic

        8            D-glucose-6-phosphate

        9            Phosphate (NADP)

    10            Magnesium chloride (MgCl)

    11            Potassium chloride (KCl)

    12            Glucose-5-phosphate dehydrogenase

    13            Liver S-9 fraction

    14            Penicillin

    15            Streptomycin

    16            Cyclophosphamide

    17            DMSO

    18            KCl

    19            Glacial acetic acid

    20            Gurr’s Giemsa Improved 266

    21            Colchicine

    22            5-bromo-2-deoxyuridine

 

Glassware’s/General requirement

 

        1            Heparinised tube (with sodium)

        2            Slides

        3            Coverslips

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

        5            Test tube racks

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

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

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

        9            Pipet tips

    10            Dispensers for buffer and S-9 mix to the test tubes

    11            Cryogenic tubes for freezing down permanent and working cultures

    12            Sterile glycerol

    13            Magnetic stir bars

    14            Mammalian tissue homogenate (S-9 fraction)

    15            Dimethylsulfoxide and other solvents

    16            Positive control chemicals

    17            Gloves

    18            Gowns

    19            Goggles or protective eyewear Biohazard bags

    20            Protective covers for the workspace

 

Equipment

        1            Autoclave

        2            Microscope (1000 X magnification)

        3            Ultraviolet light 366 nm

        4            Incubator (37 - 40°C)

        5            Gas line for keeping aseptic techniques while inoculating cultures

        6            Magnetic stirrers

        7            Balances

        8            Water purification system to generate distilled water

        9            Spectrophotometer

    10            Centrifuge (up to 10000 rpm)

    11            Revco freezer or liquid nitrogen tank

    12            Refrigerator/freezer

    13            Chemical/biological safety hood

    14            Liquid waste disposal

    15            Vacuum pump

    16            Solid waste disposal

 

Reagents and culture media

 

        1            RPMI 1600 medium (Supplied with 25 mM HEPES buffer & glutaMAX)

        2            Foetal Calf serum (FCS 10-20 %)

        3            Reagent grade PHA (2%) Phytohaemagglutamine

        4            S-9 fraction (prepared from the liver of SD rats)

        5            S-9 mix (with glucose-6-phosphate)

        6            MacIlvaine’s buffer

        7            Hoechst-33258

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