Sam:LabNotes/Microbione/2009-3-6

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Revision as of 23:41, 6 March 2009 by >Sam Chiang
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=Bacteria labeling using SYBR gree

Objective


Samples & Materials

  • Bacteria: E.coli suspension in PBS (OD600=0.913 -> around 10^9 CFU/mL = 10^6 CFU/uL)
  • SYBR 2X: Diluted from SYBR 50X stock solution


Procedures

  • Dilute SYBR 50X to 2X with RNAse-free H2O: 40 uL of 50X SYBR + 960 uL H2O -> 2X SYBR, 1000 uL.
  • Prepare four 1.5-mL tube, labeled and wrapped with foil.
  • Set up as following exp design:
                   No-treat 100uL 200uL 400uL

H2O 400 0 0 0 uL 2X SYBR 0 100 200 400 uL E. colu suspension 1 1 1 1 uL


  • Mix by gently pipetting (p1000 pipettor). Incubate at dark place for 15 min. Agitate by vortexing for every 3~4 min.
  • Prepare two