Difference between revisions of "Part:BBa K1033916:Experience"

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<partinfo>BBa_K1033916 AddReview number</partinfo>
 
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Enter the review inofrmation here.
 
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<partinfo>BBa_K1033916 AddReview 5</partinfo>
 
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<I>Hong Kong-CUHK iGEM 2017 </I>
 
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<b> Fluorescent properties of amajLime </b>
 
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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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[[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.]]
 
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<b> Charaterization of pH stability of amajLime </b>
 
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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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[[File:Amaj.PNG |center|thumb|350px|''<b>Fig.2</b>  Vary pH attributed to different fluorescent intensity of amajLime.]]
 
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    <td><b>Measurement Type</b></td>
 
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    <td>Fluorescence</td>
 
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  <tr>
 
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    <td><b>Microplate name</b></td>
 
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    <td>COSTAR 96</td>
 
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    <td><b>Scan mode</b></td>
 
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    <td>orbital averaging</td>
 
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    <td><b>Scan diameter [nm]</b></td>
 
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    <td>3</td>
 
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    <td><b>Excitation</b></td>
 
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    <td>470-15</td>
 
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  </tr>
 
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    <tr>
 
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    <td><b>Emission</b></td>
 
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    <td>515-20</td>
 
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  </tr>
 
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    <td><b>Dichronic filter </b></td>
 
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    <td>auto 491.2</td>
 
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    <td><b>Gain </b></td>
 
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    <td>500</td>
 
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      <tr>
 
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    <td><b>Focal height [nm]</b></td>
 
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    <td>9</td>
 
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<caption><p align="justify"><b>Table 1</b> Plate reader setting of fluorescent measurement</p></caption>
 
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===User Reviews===
 
===User Reviews===
 
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Latest revision as of 06:41, 21 October 2019


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Please enter how you used this part and how it worked out.

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
;