List of General Requirements for Invitro Mammalian Chromosome Aberration Test

 

Invitro Mammalian Chromosome Aberration Test

Requirement:

Chemicals:

       1          Distilled water

       2          Magnesium sulphate (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          Colchine

    22          5-bromo-2-deoxyuridine

 

Glassware’s/General requirement

 

       1          Heparinised tubes (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 eye wear Biohazard bags

    20          Protective covers for work space

 

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 liver of SD rats)

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

       6          MacIlvaine’s buffer

       7          Hoechst-33258

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