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

 
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{|width='80%' style='border:1px solid gray'
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|-
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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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|width='60%' valign='top'|
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<b> Fluorescent properties of amajLime </b>
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<p> Instead of chromorptein, we proposed 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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Fig.1 Emission and Excitation spectra
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<br>
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<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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Fig.2  Vary pH attributed to different fluorescent intensity of RFP.
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<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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    <th>Basic Settings</th>
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    <th></th>
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  <tr>
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    <td>Measurement Type</td>
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    <td>Fluorescence</td>
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  </tr>
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  <tr>
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    <td>Microplate name</td>
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    <td>COSTAR 96</td>
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  </tr>
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  <tr>
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    <td>Scan mode</td>
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    <td>orbital averaging</td>
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  </tr>
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  <tr>
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    <td>Scan diameter [nm]</td>
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    <td>3</td>
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  </tr>
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  <tr>
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    <td>Excitation</td>
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    <td>470-15</td>
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 +
  </tr>
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    <tr>
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    <td>Emission</td>
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    <td>515-20</td>
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  </tr>
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  <tr>
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    <td>Dichronic filter </td>
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    <td>auto 491.2</td>
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  </tr>
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    <tr>
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    <td>Gain </td>
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    <td>500</td>
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  </tr>
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      <tr>
 +
    <td>Focal height [nm]</td>
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    <td>9</td>
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  </tr>
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</table>
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 +
|};

Revision as of 14:38, 26 August 2017


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

Applications of BBa_K1033916

User Reviews

UNIQ422f74e82c071789-partinfo-00000000-QINU UNIQ422f74e82c071789-partinfo-00000001-QINU

•••••

Hong Kong-CUHK iGEM 2017

Fluorescent properties of amajLime

Instead of chromorptein, we proposed 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.

Fig.1 Emission and Excitation spectra
Charaterization of pH stability of amajLime

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

Fig.2 Vary pH attributed to different fluorescent intensity of RFP.

Table 1 Plate reader setting of fluorescent measurement
Basic Settings
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
;