Making TSS broth(for final volume 10 mL): Make fresh!
·10% (w/v) Polyethylene Glycol (PEG) 3350* – on media shelf; add 1 g
·50 mM Mg2+ (MgSO4 or MgCl2) – use 1M stock of MgSO4 used for NGM; add 500 uL
·Sterilise-filter into new tube
·5% (v/v) DMSO –chemical metal chest; add 500 uL
Making TSS enhanced buffer (for final volume of 10 mL):
·Add 8.2 mL of dH2O to 30 mL tube
·100mM KCl – (to prepare 1M stock: 1.491 g in 20 mL of H2O); add 1 mL
·30mM CaCl2- use 1M stock for NGM; add 0.3 mL
·50mM MgSO4 - use 1M stock for NGM; add 0.5 mL
·Vortex well and filter-sterilise into 2 mL tubes
*There are other molecular weights of PEG, so look at label carefully and choose one with 3350 on it!