IGEM:UC Berkeley/2006/KCM cells

TSS(100mL of LB): In LB  10% PEG-3350 (10g) 5% DMSO (5mL) 20mM MgCl2 (2mL from a 1M stock) pH 6.5 filter sterilize
 * 1) Overnight culture in 10 mL LB, fresh colony.
 * 2) Dilute 1:100 in 500 mL LB, grow to OD .4-.5, takes between 2-3 hours. Shake on ice to stop the growth before spin.
 * 3) Spin in prechilled bottle at 3500rpm for 15min.
 * 4) Do the rest in the cold room or on ice bucket: resuspend in 25-50mL ICE-COLD TSS with prechilled pippette.


 * 1) Aliquot into 200uL tube using multipipette.
 * 2) Freeze cells in liquid nitrogen bath.

Prechill rotor 25-50mL pipette for cell suspension 500mL centrifuge bottle whole beaker of eppendorf tubes 5mL multipipette TSS solution