Team:Korea U Seoul/Project/sub proc result

From 2013.igem.org

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                 <h1>Method</h1>
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                 <h2>Method</h2>
                 The overall reaction for precipitation is<br />
                 The overall reaction for precipitation is<br />
                 <center>Ca2+ + HCO3 - → CaCO3 + H+</center>
                 <center>Ca2+ + HCO3 - → CaCO3 + H+</center>
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                 The effect of NA and R on the rate of precipitation of CaCO3 was determined by recording the decrease in pH of a 20mM Tris HCl pH8.0 buffer solution at 10°C
                 The effect of NA and R on the rate of precipitation of CaCO3 was determined by recording the decrease in pH of a 20mM Tris HCl pH8.0 buffer solution at 10°C
                  
                  
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                 <h1>Reagents</h1>
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                 <h2>Reagents</h2>
                 <ul>
                 <ul>
                     <li>-20mM Tris-HCl buffer, pH 8.0. </li>
                     <li>-20mM Tris-HCl buffer, pH 8.0. </li>
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                 </ul>
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                 <h1>Enzyme</h1>
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                 <h2>Enzyme</h2>
                 Using Protino® Ni-NTA Columns for His-tag protein purification, purify the enzymes.<br /><br />
                 Using Protino® Ni-NTA Columns for His-tag protein purification, purify the enzymes.<br /><br />
                 These enzymes is diluted in 20mM Tris-HCl buffer, pH 8.0 + 10mM Immidazole at 4°C.
                 These enzymes is diluted in 20mM Tris-HCl buffer, pH 8.0 + 10mM Immidazole at 4°C.
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                 <h1>Procedure</h1>
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                 <h2>Procedure</h2>
                 <b>- Blank Determination</b>
                 <b>- Blank Determination</b>
                 Add 0.5 ml of 1M CaCl2 to a 15 ml tube cotaining 0.5 ml of 1M NaHCO3 and 9 ml of 20mM Tris-HCl buffer. Immediately start a stop watch and record the time and pH up to 20 minutes.
                 Add 0.5 ml of 1M CaCl2 to a 15 ml tube cotaining 0.5 ml of 1M NaHCO3 and 9 ml of 20mM Tris-HCl buffer. Immediately start a stop watch and record the time and pH up to 20 minutes.

Revision as of 21:08, 27 September 2013

CaCO3 Precipitation Assay

Method

The overall reaction for precipitation is
Ca2+ + HCO3 - → CaCO3 + H+
According to that equation, CaCO3 generated the more the pH will be lowered.

The effect of NA and R on the rate of precipitation of CaCO3 was determined by recording the decrease in pH of a 20mM Tris HCl pH8.0 buffer solution at 10°C

Reagents

  • -20mM Tris-HCl buffer, pH 8.0.
  • -20mM Tris-HCl buffer, pH 8.0 + 1M Imidazole.
  • -1M NaHCO3.
  • -1M CaCl2.

Enzyme

Using Protino® Ni-NTA Columns for His-tag protein purification, purify the enzymes.

These enzymes is diluted in 20mM Tris-HCl buffer, pH 8.0 + 10mM Immidazole at 4°C.

Procedure

- Blank Determination Add 0.5 ml of 1M CaCl2 to a 15 ml tube cotaining 0.5 ml of 1M NaHCO3 and 9 ml of 20mM Tris-HCl buffer. Immediately start a stop watch and record the time and pH up to 20 minutes. - Enzyme Determination Add freshly diluted enzyme to a 15 ml tube cotaining 0.5 ml of 1M NaHCO3 and 9 ml of 20mM Tris-HCl buffer. Add Add 0.5 ml of 1M CaCl2. Immediately start a stop watch and record the time and pH up to 20 minutes.

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