Biotechnology Procedures and Experiments Handbook
By S. Harisha

Appendix C: Reagents Required for Tissue Culture Experiments

STOCK SOLUTIONS AND MEDIA

Sterile water bottles

200 mL, 500 mL, or 950 mL water.

Autoclave.

0.05 M CaCl2 (per 200 mL)

CaCl 2 2H 2O

1.5 g

Add 198 mL water.

Autoclave.

0.5 M CaCl2 (per 200 mL)

CaCl 2 2H 2O

15.0 g

Add 188 mL water.

Autoclave.

0.1 M MgSO 4 (per 200 mL)

MgSO 4 (anhydrous)

2.4 g

Add 200 mL water.

Autoclave.

10% MSG (for 200 mL)

Glutamic acid (Na)

20.0 g

Add 188 mL water.

Autoclave.

40% Glucose (per 200 mL)

Glucose(dextrose)

80.0 g

Add water to 200 mL

(~145 150 mL).

Autoclave.

10% CAA (per 200 mL)

Casein acid hydrolysate

20.0 g

Add 186 mL water.

Autoclave.

10% Yeast (per 200 mL)

Yeast extract

20.0 g

Add 150 160 mL water. Autoclave.

Add 150 L ?-mercaptoethanol after autoclaving.

20X Tryptone/NaCl (per 200 mL)

Tryptone

40.0 g

NaCl

20.0 g

Add 190 mL water. Autoclave.

Add 150 L ?-mercaptoethanol after autoclaving.

10X M9 stock (for 200 mL)

Na 2HPO 4 (anhydr.)

12.0 g

KH 2PO 4

6.0 g

NaCl

1.0 g

NH 4Cl

2.0 g

Add 196 mL H 2O. Autoclave.

Note

Leave out NH 4Cl to prepare 10X M9-N.

10X M9-N stock (for 200 mL)

Na 2HPO 4 (anhydr.)

12.0 g

KH 2PO 4

6.0 g

NaCl

1.0 g

Add 196 mL H 2O. Autoclave.

Note

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Copyright Infinity Science Press LLC 2007 under license agreement with Books24x7
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