User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2013/02/06

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** UV-vis spectrometer was set under photometric method, with wavelength set at 650nm.
** UV-vis spectrometer was set under photometric method, with wavelength set at 650nm.
** A distilled water sample placed in plastic cuvette was placed into the spectrometer to eliminate background noise.
** A distilled water sample placed in plastic cuvette was placed into the spectrometer to eliminate background noise.
-
** 2mL of DH10B and DH5α-T1 cells were individually placed into the spectrometer to measure absorbance.
+
** 2mL of DH10B and DH5α-T1 cells were individually placed into a plastic cuvette into the spectrometer to measure absorbance.
** The absorbance for each cell after 15 hours of growth is shown in table below:
** The absorbance for each cell after 15 hours of growth is shown in table below:
-
 
+
{| {{table}}
-
* For future procedure on clay-incorporated silver, please refer to [[Melissa's Notebook]]
+
| align="center" style="background:#f0f0f0;"|'''Cells'''
 +
| align="center" style="background:#f0f0f0;"|'''Absorbance'''
 +
|-
 +
| DH10B||1.82
 +
|-
 +
| DH5α-T1||1.8
 +
|-
 +
|
 +
|}
 +
* For future procedure on clay-incorporated silver, please refer to [[User:Melissa Novy/Notebook/CHEM-572/2013/02/06
 +
|Melissa's Notebook]]
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Revision as of 10:05, 8 February 2013

Experimental Biological Chemistry II Main project page
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Purpose

  • Remake LB+agar plates
  • Make glycerol stock of DH10B and DH5α-T1 cells
  • Test absorbance of cells in culture grew last night for cell concentration

Procedure

  • Because agar plates made on 2013/01/31 liquified, new plates were made.
    • 12.5g of LB media powder were weighted and placed in a 750mL Erlenmeyer flask.
    • 7.5g of Agar were weighted out and placed in the same 750mL Erlenmeyer flask.
    • 500mL distilled water was measured out and placed in the same 750mL Erlenmeyer flask.
    • The solution was autoclaved per liquid cycle.
    • After autoclave, the agar were placed in room temperature to cool down to 60°C.
    • Approximately 25mL of agar were poured into each petri dish.
    • The agar plates were left in room temperature under flame to solidify.
    • The agar plates were stored in fidge for future use.
  • Glycerol stock of the two cell lines: DH10B and DH5α-T1 cells were made for future use.
    • Two 5mL Gyro® Tubes were obtained, one for DH10B cells and another for DH5α-T1 cells.
    • 150mL of autoclaved 60% glycerol in water was pipetted into the Gyro® Tubes prior.
    • 850mL of DH10B cells, and 850mL of DH5α-T1 cells were pipetted into each of Gyro® Tubes, respectively.
    • Each cell culture containing the cell and glycerol were vortexed until mix.
    • Cells were placed in -80°C for storage.
  • Absorbance of cells were taken using UV-vis spectroscopy to measure the concentration of cells that had been grown over night.
    • UV-vis spectrometer was set under photometric method, with wavelength set at 650nm.
    • A distilled water sample placed in plastic cuvette was placed into the spectrometer to eliminate background noise.
    • 2mL of DH10B and DH5α-T1 cells were individually placed into a plastic cuvette into the spectrometer to measure absorbance.
    • The absorbance for each cell after 15 hours of growth is shown in table below:
Cells Absorbance
DH10B1.82
DH5α-T11.8
  • For future procedure on clay-incorporated silver, please refer to [[User:Melissa Novy/Notebook/CHEM-572/2013/02/06
Melissa's Notebook]]


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