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APPENDIX

Appendix 1: Composition of Nutrient Agar composition

S. No Ingredients Quantity ( g/l )

1 Peptic digest of animal tissue 5

2 Yeast extract 1.5

3 Beef extract 1.5

4 Sodium chloride 5

5 Agar 15

Preparation of nutrient agar


28 grams of nutrient agar was dissolved in 1000 ml distilled water. The mixture heated to Boil to

dissolve medium completely .Media was sterilized completely by autoclaving at 121 for minutes.
Appendix 2: Composition of MacConkey Agar

S. no Ingredients Quantity ( g/l )

1 Lactose monohydrate 10

2 Bile salt 1.5

3 Sodium chloride 5

4 Natural red 0.03

5 Peptone (Pancreatic digest of gelatin) 17

6 Proteose Peptone (meat and casein) 3

7 Agar 13.5

Preparation of McConkey’s agar

50 g of the media was mixed in 1000ml distilled water .Heat with frequent agitation and boil for

one minute to completely dissolve the medium. Autoclave at 121 for 15 minutes.
Appendix 03 Composition of Manitol salt agar (MSA)

S. No Ingredients Quantity(gm/l)

1 Beef extract 1.0

2 Phenol red 0.025

3 D-Manitol 10.0

4 Peptic digest of animal tissue 5.0

5 Pancreatic digest of casein 5.0

6 Sodium chloride 75.0

6 Agar 15.0

Suspend 111.025gm of MSA in 1000ml water and autoclave it at 121°C for 15 mints.
Appendix 04 Composition of phosphate buffer saline

S. No Ingredients Quantity (g/ml)

1 Sodium chloride 8

2 Potassium chloride 0.2

3 Sodium hydrogen phosphate 1.44

4 Potassium hydrogen phosphate 0.24

Preparation of phosphate buffer saline


Mix all ingredients in distilled water, adjust the PH to 7.3± 0.2 with the help of PH meter by 1M
hydrochloric acid and 1M hydro oxide, added in 10 ml and autoclaved for 15 minutes at 121°C

S.No Ingredients Quantity g/l

1 Nacl 1

2 yast extract 0.5

3 Tryptone 1

4 Bacto agar 1.5