∞ generated and posted on 2015.05.08 ∞
With refrigeration, phage stocks can last for decades in this buffer.
| Dilutent and Respension Buffer (TSG) for phages can consist of Tris-HCl (pH 7.4-7.5), 0.01 M; NaCl, 0.15 M; Gelatin, 0.03%. Autoclave ingredients together. |
The following is the recipe adjusted for various volumes:
| Volume: | 0.5 L | 1 L | 1.5 L | 2 L | 5 L | 10 L | 20 L |
|---|---|---|---|---|---|---|---|
| Tris-HCl | 0.8 g | 1.6 g | 2.4 g | 3.2 g | 7.9 g | 15.8 g | 31.5 g |
| NaCl | 4.4 g | 8.8 g | 13.1 g | 17.5 g | 43.8 g | 87.7 g | 175.3 g |
| Gelatin | 0.15 g | 0.3 g | 0.45 g | 0.6 g | 1.5 g | 3.0 g | 6.0 g |
Adjust pH to 7.4 to 7.5; Autoclave.
For long-term storage it is recommended that you first purify your phage stock such as via membrane filtration (e.g., 0.22 μm) and/or following high-speed centrifugation, pelleting, and then gradual (e.g., overnight) pellet resuspension.
For most phages, you can also place a modest amount of chloroform to prevent growth of contaminants, e.g., 0.5 ml into screw capped tubes for storage such as of 10 ml of phage stock.
I tend to store unused buffer in 100-ml aliquots.
Recipe along with phage storage instructions are presented by Carlson and Miller, 1994: Carlson, K., Miller, E. S. (1994). Working with T4. in The Molecular Biology of Bacteriophage T4, Karam, J.D. (ed.), American Society for Microbiology Press, Washington D.C., pp. 421-426.