McClean: Bacterial Glycerol 65%

Overview

Use this solution for freezing down E. coli bacterial stocks. The desired final solution is:

• 65% glycerol (vol/vol)
• 0.1 M MgSO4
• 0.025 M Tris-Cl pH 8

Materials

1. Glycerol (Fisher BP229)
2. MgSO4 1M Stock Solution (Fisher M65; 120.366g/L)
3. 1M Tris-HCl, pH'd with NaOH to pH 8

Procedure

Combine:

• 52 ml glycerol
• 8 ml 1M MgSO4 solution
• 2 ml 1M Tris-HCl

Add sterile H₂O to 80 ml total volume. Autoclave on 30 min liquid cycle.

Notes

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Contact

• Megan N McClean 14:01, 20 July 2011 (EDT)

or instead, discuss this protocol.