TRYPTIC SOY BROTH w/ 15% GLYCEROL INTENDED USE Remel Tryptic Soy Broth w/ 15% Glycerol is a liquid medium recommended for use in long-term storage of bacteria. SUMMARY AND EXPLANATION This medium is recommended in Clinical Microbiology Procedures Handbook and by Clinical and Laboratory Standards Institute for the maintenance of stock cultures.1,2 When inoculated into Tryptic Soy Broth w/ 15% Glycerol and frozen at or below -40°C, suspensions of bacteria remain viable for several months. PRINCIPLE Casein and soy peptones provide amino acids and nitrogenous compounds for the growth of bacteria. Sodium chloride is a source of essential electrolytes and maintains osmotic equilibrium. Dipotassium phosphate is a pH buffer. Dextrose is a carbon energy source. Glycerol is a cryoprotective agent that provides intracellular and extracellular protection against freezing and promotes long-term preservation. REAGENTS (CLASSICAL FORMULA)* Casein Peptone........................................................... 17.0 g Dipotassium Phosphate ................................................2.5 g Sodium Chloride............................................................ 5.0 g Dextrose ........................................................................2.5 g Soy Peptone.................................................................. 3.0 g Glycerol .....................................................................150.0 ml Demineralized Water.................................................850.0 ml pH 7.3 ± 0.2 @ 25°C *Adjusted as required to meet performance standards. PROCEDURE 1. Inoculate the medium with a pure culture using a sterile inoculating loop or swab. 2. Place the tube with cap tightened at or below -40°C. 3. Organisms that have been inoculated and frozen in this medium should be thawed and checked for viability at appropriate intervals, following established laboratory guidelines. QUALITY CONTROL All lot numbers of Tryptic Soy Broth w/ 15% Glycerol have been tested using the following quality control organisms and have been found to be acceptable. Testing of control organisms should be performed in accordance with established laboratory quality control procedures. If aberrant quality control results are noted, patient results should not be reported. CONTROL INCUBATION RESULTS Escherichia coli ATCC® 25922 Aerobic, 24 h @ -70°C Growth on subculture Pseudomonas aeruginosa ATCC® 27853 Aerobic, 24 h @ -70°C Growth on subculture Staphylococcus aureus ATCC® 25923 Aerobic, 24 h @ -70°C Growth on subculture LIMITATIONS 1. Growth on subculture of isolates preserved in Tryptic Soy Broth w/ 15% Glycerol may depend on inoculum size, duration of storage, and the fastidious nature of the organism. BIBLIOGRAPHY 1. Isenberg, H.D. 2004. Clinical Microbiology Procedures Handbook. 2nd ed. ASM Press, Washington, D.C. 2. Clinical and Laboratory Standards Institute (CLSI). 2009. Performance Standards for Antimicrobial Disk Susceptibility Tests; Approved Standard, 10th ed. M2-A10. CLSI, Wayne, PA. Refer to the front of Remel Technical Manual of Microbiological Media for General Information regarding precautions, product storage and deterioration, specimen collection, storage and transportation, materials required, quality control, and limitations. ATCC is a registered trademark of American Type Culture Collection. IFU 65024, Revised February 1, 2010 Printed in U.S.A. 12076 Santa Fe Drive, Lenexa, KS 66215, USA General Information: (800) 255-6730 Website: www.remel.com Email: [email protected] Local/International Phone: (913) 888-0939 International Fax: (913) 895-4128 .