Team:Korea U Seoul/Project/sub proc result

From 2013.igem.org

(Difference between revisions)
Line 1: Line 1:
<html>
<html>
-
    <head>
 
-
        <style>
 
-
            h2 { margin-bottom: 30px}
 
-
        </style>
 
-
    </head>
 
-
    <body style="">
 
-
        <div class="content_wrapper" style="">
 
-
            <div class="title">
 
-
                <div class="title_">
 
-
                  CaCO3 Precipitation Assay
 
-
                </div>
 
-
                <div class="left_line"></div><div class="right_line"></div>
 
-
            </div>            a<html>
 
     <head>
     <head>
         <style>
         <style>
Line 45: Line 32:
                 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.<br /><br /><br />
                 These enzymes is diluted in 20mM Tris-HCl buffer, pH 8.0 + 10mM Immidazole at 4°C.<br /><br /><br />
-
 
-
                <h2>Procedure</h2>
 
-
                <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.
 
-
                <b>- Enzyme Determination</b>
 
-
                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.
 
-
 
-
            </p>
 
-
            </div>
 
-
                    </div>
 
-
    </body>
 
-
</html>
 
-
 
-
            <div class="content_">
 
-
            <p>
 
-
                <h2>Method</h2>
 
-
                The overall reaction for precipitation is<br />
 
-
                <center>Ca2+ + HCO3 - → CaCO3 + H+</center>
 
-
                According to that equation, CaCO3 generated the more the pH will be lowered.<br /><br />
 
-
                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
 
-
               
 
-
                <h2>Reagents</h2>
 
-
                <ul>
 
-
                    <li>-20mM Tris-HCl buffer, pH 8.0. </li>
 
-
                    <li>-20mM Tris-HCl buffer, pH 8.0 + 1M Imidazole.</li>
 
-
                    <li>-1M NaHCO3.</li>
 
-
                    <li>-1M CaCl2.</li>
 
-
                </ul>
 
-
               
 
-
                <h2>Enzyme</h2>
 
-
                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.
 
                 <h2>Procedure</h2>
                 <h2>Procedure</h2>

Revision as of 21:10, 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.

Retrieved from "http://2013.igem.org/Team:Korea_U_Seoul/Project/sub_proc_result"