MIT Synthetic Biology Working Group Standardized FACS Protocol Draft 1 – 9/20/04

This protocol sets out to standardize the components of a FACS experiment necessary to allow repeatable and comparable results. The output of these experiments is typically fluorescence from a genetic regulatory network.

Media and Glassware

Provisionally, supplemented M9 media will be used for all cultures. The media recipe used by CC and JCB is –

• 1x M9 Minimal Salts (from Sigma):
• 12.8 g/L NaHPO4.7H20
• 3.0 g/L KH2PO4
• 0.5 g/L NaCl
• 1.0 g/L NH4Cl
• 0.1mM MgSO4
• 2mM CaCl2
• 0.4% Glucose/Glycerol
• 0.1% Casamino Acids