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<br>MATERIALS <br>- SOB/LB Media <br>- Glycerol (100%) <br>- Sterile Milli-Q dH2O <br>- Liquid nitrogen <br>CHECK-LIST PROCEDURE <br>- Choose colony put into 5 mL of LB medium in 15 mL falcon. <br>- Incubate at 37 C @ 240 rpm not longer than 12-14 h. <br>- Centrifuge 5000 rpm 5min. <br>- Discard 4 mL solution from falcon. <br>- Add 176 uL glycerol into falcon to 15% final concentration. <br>- Vortex! <br>- Aliquot 150 uL samples. <br>- Quick-freeze in liquid nitrogen <br>- Place in -80 C deep freeze

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