Super Broth No. II

Super Broth No. II is used for the cultivation of recombinant strains of Escherichia coli

Composition**

Final pH ( at 25°C) 7.2±0.2

**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 49.1 grams in 1000 ml distilled water containing 5 ml glycerol. Heat if necessary to dissolve the medium completely. Dispense in tubes. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 mins.

Principle And Interpretation

The rearrangement of genetic information within and among DNA molecule encompasses a variety of processes, collectively placed under the heading of genetic recombination (1). Bacteria that have undergone recombination, need to be grown in an enriched medium. Super Broth No. II is a modification of Super Broth, developed by Tartoff and Hobbs (2).

High amount of tryptone and yeast extract, make the medium highly nutritive for the growth of recombinant strains of E.coli. Phosphate provide buffering to the medium.

Quality Control

Appearance: Cream to yellow coloured homogeneous free flowing powder

Colour and Clarity of prepared medium: Amber coloured clear solution without any precipitate

Reaction: Reaction of 4.9% w/v aqueous solution at 25°C. pH : 7.2±0.2

pH: 7.00-7.40

Cultural Response: M1689: Cultural characteristics observed after an incubation at 35-37°C for 18- 24 hours.

Storage and Shelf Life

Store below 30°C in tightly closed container and the prepared medium at 2-8°C. Use before expiry date on the label.

Reference

1. Nelson D.L, Cox M.M, 2005 Lehninger Principles of Biochemistry, 4th edi, W.H. Freeman and Company. New York.
2. Tartoff and Hobbs . 1987. Bethesda Research Laboratories FOCUS 9:12.