Ingredients

• PEG 3350 (5g)
• DMSO (2.5mL)
• MgCl2 * 6H2O (0.3g)
• LB Broth (Miller Mod)

Equipment

• Analytical Balance accurate to 0.001g
• 50mL STERILE Centrifuge Tube
• 0.22um Syringe Filter (25mm or larger diameter for easier press)
• 60mL Luer Lock Syringe

Step By Step

1. To a sterile 50mL centrifuge tube add 5 grams of PEG 3350 (Miralax).
2. Then add either 0.3g of MgCl2*6H2O or 1.5mL of 1M MgCl2.
3. Next add 2.5mL of pure DMSO.
4. Finally, add sterile LB Broth (Miller Mod) to the 40mL mark, vortex until fully dissolved, and top off to the 50mL mark.
5. Filter sterilize media with a 0.22um syringe filter and store at -20C.

Notes

References

Miller, J.H. 1972. Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor.N.Y.