Difference between revisions of "Part:BBa K1033916:Experience"
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===Applications of BBa_K1033916===
 
===Applications of BBa_K1033916===
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===User Reviews===
 
===User Reviews===
 
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<b> Fluorescent properties of amajLime </b>
 
<b> Fluorescent properties of amajLime </b>
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<p> Instead of chromorptein, we proposed that amajLime is a fluorescent protein with the property of showing pigmentation, like BBa_E1010. We charaterised spectral properties of amajLime and found its max excitation and emission wavelength at 445 nm nd 485 nm respectively.</p>
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<p> Although amajLime is described as chromoprotein in main page, we characterised its spectral properties and found the max excitation and emission wavelength at 445 nm nd 485 nm respectively.</p>
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Fig.1 Emission and Excitation spectra
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[[File:Ex-Em. amajlime.png|width='60%' valign='top'| |center|thumb|550px|''<b>Fig.1</b>  Emission and Excitation spectra in blue and green respectively.]]
 
<br>
 
<br>
 
<b> Charaterization of pH stability of amajLime </b>
 
<b> Charaterization of pH stability of amajLime </b>
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<p>We grew C41 bacteria with parts BBa_J61002 with constituitive promoter: J23100, in 2XYT for 24 hours. Purifying the amajLime by Ion Exchange Chromatography and Hydrophobic Interaction Chromatography, we measured the fluoresece of purified amajLime, which is diluted to 10µg/100µl (total 200µl) in triplicates, in different buffers (ranges from pH2 to pH12). The result shows that the stability drops dramatically in condition below 6 and attains maxima fluorescence at pH 7</p>
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<p>We transformed part BBa_K1033916 with constituitive promoter: J23100 in C41 and grew in 2XYT for 24 hours. Purifying the amajLime by Ion Exchange Chromatography and Hydrophobic Interaction Chromatography, we measured the fluoresece of purified amajLime, which is diluted to 10µg/100µl (total 200µl) in triplicates, in different buffers (ranges from pH2 to pH12). The result shows that the stability drops dramatically in pH condition below 6 and attains maxima fluorescence at pH 8.</p>
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Fig.2  Vary pH attributed to different fluorescent intensity of RFP.
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 +
<div align="center">
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[[File:Amaj.PNG |center|thumb|350px|''<b>Fig.2</b> Vary pH attributed to different fluorescent intensity of amajLime.]]
 
<table cellpadding="2" border="1px" cellspacing="0" align="center" width="70%">
 
<table cellpadding="2" border="1px" cellspacing="0" align="center" width="70%">
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<caption><p align="justify"><b>Table 1</b> Plate reader setting of fluorescent measurement</p></caption>
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−
  <tr>
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    <td><b>Measurement Type</b></td>
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    <th>Basic Settings</th>
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−
    <th></th>
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−
  <tr>
+
−
    <td>Measurement Type</td>
+
 
     <td>Fluorescence</td>
 
     <td>Fluorescence</td>
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  </tr>
+
 
 
   <tr>
 
   <tr>
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     <td>Microplate name</td>
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     <td><b>Microplate name</b></td>
 
     <td>COSTAR 96</td>
 
     <td>COSTAR 96</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
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     <td>Scan mode</td>
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     <td><b>Scan mode</b></td>
 
     <td>orbital averaging</td>
 
     <td>orbital averaging</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
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     <td>Scan diameter [nm]</td>
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     <td><b>Scan diameter [nm]</b></td>
 
     <td>3</td>
 
     <td>3</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
−
     <td>Excitation</td>
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     <td><b>Excitation</b></td>
 
     <td>470-15</td>
 
     <td>470-15</td>
  
 
   </tr>
 
   </tr>
 
     <tr>
 
     <tr>
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     <td>Emission</td>
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     <td><b>Emission</b></td>
 
     <td>515-20</td>
 
     <td>515-20</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
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     <td>Dichronic filter </td>
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     <td><b>Dichronic filter </b></td>
 
     <td>auto 491.2</td>
 
     <td>auto 491.2</td>
 
   </tr>
 
   </tr>
 
     <tr>
 
     <tr>
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     <td>Gain </td>
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     <td><b>Gain </b></td>
 
     <td>500</td>
 
     <td>500</td>
 
   </tr>
 
   </tr>
 
       <tr>
 
       <tr>
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     <td>Focal height [nm]</td>
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     <td><b>Focal height [nm]</b></td>
 
     <td>9</td>
 
     <td>9</td>
 
   </tr>
 
   </tr>
 +
<caption><p align="justify"><b>Table 1</b> Plate reader setting of fluorescent measurement</p></caption>
 
</table>
 
</table>
  
 
|};
 
|};

Latest revision as of 06:41, 21 October 2019


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Applications of BBa_K1033916

User Reviews

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•••••

Hong Kong-CUHK iGEM 2017

Fluorescent properties of amajLime

Although amajLime is described as chromoprotein in main page, we characterised its spectral properties and found the max excitation and emission wavelength at 445 nm nd 485 nm respectively.

Fig.1 Emission and Excitation spectra in blue and green respectively.


Charaterization of pH stability of amajLime

We transformed part BBa_K1033916 with constituitive promoter: J23100 in C41 and grew in 2XYT for 24 hours. Purifying the amajLime by Ion Exchange Chromatography and Hydrophobic Interaction Chromatography, we measured the fluoresece of purified amajLime, which is diluted to 10µg/100µl (total 200µl) in triplicates, in different buffers (ranges from pH2 to pH12). The result shows that the stability drops dramatically in pH condition below 6 and attains maxima fluorescence at pH 8.

Fig.2 Vary pH attributed to different fluorescent intensity of amajLime.
Measurement Type Fluorescence
Microplate name COSTAR 96
Scan mode orbital averaging
Scan diameter [nm] 3
Excitation 470-15
Emission 515-20
Dichronic filter auto 491.2
Gain 500
Focal height [nm] 9

Table 1 Plate reader setting of fluorescent measurement
;