Sam:LabNotes/Microbiome-new/2009-2-23

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Plate counging for diluted E. coli used in previous MDA exp[edit]

Objective[edit]

  • To confirm the bacterai number in previous Bac MDA experiment

Procedures[edit]

  • Prepare the non-antibiotics LB plates
  • Plating
    • No.1 (1000 cell/uL) -> 1/10 dilution -> 50 uL plating on w/o antibiotics LB agar plates x 2 duplicates
    • No.2 ( 100 cell/uL) -> 1/10 dilution -> 50 uL plating on w/o antibiotics LB agar plates x 2 duplicates
    • No.3 ( 10 cell/uL) -> 1/10 dilution -> 50 uL plating on w/o antibiotics LB agar plates x 2 duplicates
  • Incubate at 37 degree incubator over night.
  • Count the plate the next day.

Results[edit]

  • Expected number of plate counting
    • No.1 1/10 diluted 1000 cell/uL x 50 uL = 5000 cell --- not countable
    • No.2 1/10 diluted 100 cell/uL x 50 uL = 500 cell --- not countable
    • No.3 1/10 diluted 10 cell/uL x 50 uL = 50 cell --- 53 cells (Replicate 1) and 79 cells (Replicates 2)
  • The results showed that OD600 = 1 -> 10^9 CUF/mL of E. coli is a good estimation.