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

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(Autocreate 2013/02/06 Entry for User:Keyun_Wang/Notebook/Experimental_Biological_Chemistry_I)
Current revision (09:07, 8 February 2013) (view source)
(Procedure)
 
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
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|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==
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==Purpose==
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* Insert content here...
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* Remake LB+agar plates
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* Make glycerol stock of DH10B and DH5α-T1 cells
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* Test absorbance of cells in culture grew last night for cell concentration
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==Procedure==
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* Because agar plates made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2013/01/30|2013/01/30]] liquified, new plates were made.
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** 12.5g of LB media powder were weighted and placed in a 750mL Erlenmeyer flask.
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** 7.5g of Agar were weighted out and placed in the same 750mL Erlenmeyer flask.
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** 500mL distilled water was measured out and placed in the same 750mL Erlenmeyer flask.
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** The solution was autoclaved per liquid cycle.
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** After autoclave, the agar were placed in room temperature to cool down to 60°C.
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** Approximately 25mL of agar were poured into each petri dish.
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** 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:
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Cells'''
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| align="center" style="background:#f0f0f0;"|'''Absorbance'''
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|-
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| DH10B||1.82
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|-
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| DH5α-T1||1.8
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|-
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|
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|}
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* 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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Current revision

Experimental Biological Chemistry II Main project page
Previous entry      Next entry

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 2013/01/30 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


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