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

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Experimental Biological Chemistry II</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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==Entry title==
==Purpose==
* Insert content here...
* 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 [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2013/01/31|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 the spectrometer to measure absorbance.
** The absorbance for each cell after 15 hours of growth is shown in table below:
* For future procedure on clay-incorporated silver, please refer to [[Melissa's Notebook]]


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Revision as of 07:02, 8 February 2013

Experimental Biological Chemistry II <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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 the spectrometer to measure absorbance.
    • The absorbance for each cell after 15 hours of growth is shown in table below:


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