Sam:LabNotes/Microbiome/2009-3-9: Difference between revisions

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>Sam Chiang
(New page: ='''E. coli labeling using SYBR Green I and YOYO-1'''= ==Objective== *To confirm the bacteria staining with revised procedures **Using FITC filter, 20X objective, No cover slip *The sta...)
 
>Sam Chiang
Line 24: Line 24:
*Mix the E. coli with either 10 uL 2X SYBR Green I or 2 uL of YOYo-1 dye, gently pipeting for mixing
*Mix the E. coli with either 10 uL 2X SYBR Green I or 2 uL of YOYo-1 dye, gently pipeting for mixing


*Leave the slide in dark box (slide storage box)
*Leave the slide in dark box (slide storage box)for incubation 15 min in room temp
 
==Results==
  [[Image:SNAP-151939-0309-sybr-20x-B&W.jpg|SNAP-151939-0309-sybr-20x-B&W]]
  [[Image:SNAP-153406-0004-sybr-20x-fitc.jpg|SNAP-153406-0004-sybr-20x-fitc]]
  [[Image:SNAP-153514-0005-sybr-20x-fitc.jpg|SNAP-153514-0005-sybr-20x-fitc]]

Revision as of 23:52, 9 March 2009

E. coli labeling using SYBR Green I and YOYO-1

Objective

  • To confirm the bacteria staining with revised procedures
    • Using FITC filter, 20X objective, No cover slip
  • The stained bacteria will be used for microwell application


Material and Samples

  • Samples: Diluted E. coli, 10^6 CFU/uL (stored in PBS, 4C)
  • YOYO®-1 iodide (491/509) - 1 mM solution in DMSO
  • Diluted SYBR Green I (diluted in H2O, stored in 4C)


Procedures

  • Drop 1 uL of E. coli on the glass slide
  • Mix the E. coli with either 10 uL 2X SYBR Green I or 2 uL of YOYo-1 dye, gently pipeting for mixing
  • Leave the slide in dark box (slide storage box)for incubation 15 min in room temp

Results

  SNAP-151939-0309-sybr-20x-B&W
  SNAP-153406-0004-sybr-20x-fitc
  SNAP-153514-0005-sybr-20x-fitc