Team:Korea U Seoul/Project/sub proc result
From 2013.igem.org
(Difference between revisions)
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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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<li>-20mM Tris-HCl buffer, pH 8.0. </li> | <li>-20mM Tris-HCl buffer, pH 8.0. </li> | ||
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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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<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 isThe 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.