Blasticidin S HCl solution (CAS 3513-03-9)

Caution

Toxic if swallowed. Contains Blasticidin S. Avoid contact with skin and eyes. Wear suitable protective clothing and gloves. In case of accident or if you feel unwell, seek medical advice immediately.



Storing Blasticidin

• Do not subject Blasticidin solution to multiple freeze-thaw cycles (do not store in a frost-free freezer).

• When stored properly at –5°C to –20°C, Blasticidin is stable for up to 9 months. Store medium containing Blasticidin at 4°C for up to 2 weeks.

• Upon thawing, use what you need and discard any unused portion.



Blasticidin Selection in E. coli

For selection of Blasticidin-resistant E. coli, use Low Salt LB medium (10 g Tryptone, 5 g sodium chloride, 5 g yeast extract) containing 50–100 µg/mL Blasticidin. Optimize the Blasticidin concentration for the bacterial strain being used. If a lawn of bacteria forms on the Low Salt LB plate instead of individual bacterial colonies, increase the Blasticidin concentration in the plates to 100 µg/mL.

Note: The salt concentration of the medium must remain low (<90 mM). Failure to lower the salt content of your LB medium inhibits Blasticidin, and prevents selection unless a higher Blasticidin concentration is used.



Blasticidin Selection in Yeast

The concentration of Blasticidin required for selection in yeast varies depending on the species, strain, and type of medium used. Use a range of 25–300 µg/mL Blasticidin for selection in yeast. Standard protocols recommend performing a kill curve for each species, strain, and medium used to determine the appropriate Blasticidin concentration to use for selecting resistant cells.



Blasticidin Selection in Mammalian Cells

The Blasticidin concentration required for selection in mammalian cells varies depending on the cell line used. Use 2–10 µg/mL Blasticidin for selection in mammalian cells. Standard protocols recommend performing a kill curve as described below to determine the appropriate Blasticidin concentration to use for selecting resistant cells.



Determining blasticidin sensitivity

1. Plate cells at approximately 25% confluency. Prepare a set of 6 plates. Allow cells to adhere overnight.

2. The next day, substitute culture medium with medium containing varying concentrations of Blasticidin (e.g. 0, 2, 4, 6, 8, 10 µg/mL Blasticidin).

3. Replenish the selective media every 3–4 days, and observe the percentage of surviving cells.

4. Determine the appropriate concentration of Blasticidin that kills the cells within 10–14 days after addition of the antibiotic.