Difference between revisions of "Part:BBa B0030:Experience"

 
 
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__NOTOC__
 
__NOTOC__
 
This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
how you used this part and how it worked out.
 
how you used this part and how it worked out.
  
 +
===Characterizations of BBa_B0030===
 +
The Alverno_Ca team characterized BBa_K2066046, which includes BBa_B0030, as well as a series of other plasmids using TX-TL, a mean of in vitro transcription and translation using cell extract. Below is a graph of normalized expression of BBa_K2066046 in comparison to the other palsmids in the sequence.
 +
https://static.igem.org/mediawiki/2016/4/4f/T--Alverno_CA--W%26M_corrected.png
 +
 +
This graph shows the fluorescence of the plasmids, over a period of 12 and a half hours, in a plate reader.
 +
https://static.igem.org/mediawiki/2016/c/c1/T--Alverno_CA--W%26M_plasmid_time_traces_in_TXTL%282%29.png"
 
===Applications of BBa_B0030===
 
===Applications of BBa_B0030===
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 +
William and Mary iGEM 2016
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 +
https://static.igem.org/mediawiki/2016/5/58/T--William_and_Mary--RBS_Characterization_Curves.png
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 +
We characterized the strengths of the Anderson RBS library.
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===User Reviews===
 
===User Reviews===
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This review comes from the old result system and indicates that this part worked in some test.
 
This review comes from the old result system and indicates that this part worked in some test.
 
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{|width='80%' style='border:1px solid gray'
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|-
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|width='10%'|
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<partinfo>BBa_R0051 AddReview 3</partinfo>
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<I> Aberdeen_Scotland 2009 </I>
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|width='60%' valign='top'|
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The miniprep, single and double digests all worked. However we did not use this part for further cloning.
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|}
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<!-- DON'T DELETE --><partinfo>BBa_B0030 EndReviews</partinfo>
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<!-- DON'T DELETE --><partinfo>BBa_B0030 StartReviews</partinfo>
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<!-- Template for a user review
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{|width='80%' style='border:1px solid gray'
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|-
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|width='10%'|
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<partinfo>BBa_B0030 AddReview 4</partinfo>
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<I>Username</I>
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|width='60%' valign='top'|
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Enter the review inofrmation here.
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|};
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<!-- End of the user review template -->
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{|width='80%' style='border:1px solid gray'
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|-
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|width='10%'|
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<partinfo>BBa_B0030 AddReview 5</partinfo>
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<I>UNIPV-Pavia iGEM 2011</I>
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|width='60%' valign='top'|
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<html>
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<em>
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NB: unless differently specified, all tests were performed in <a href='http://2011.igem.org/Team:UNIPV-Pavia/Protocols#MG1655Z1'><em>E. coli</em> MGZ1</a> in M9 supplemented medium at 37°C in low copy plasmid <A HREF="https://parts.igem.org/wiki/index.php/Part:pSB4C5">pSB4C5</a>.
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</em>
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<br><br>
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See <a href="https://parts.igem.org/Part:BBa_B0034:Experience">Experience page of BBa_B0034</a> for more details.
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<br><br>
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Estimated efficiencies in pSB4C5 plasmid with -RBSx-mRFP-TT coding sequence under the control of the specified promoter:
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<br><br>
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<div align="center"><table class='data' width='70%' border='1'><tr>
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<td class='row'><b>RBS</b></td>
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<td class='row'><b>eff<sub>p<sub>Lux</sub></sub></b></td>
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<td class='row'><b>eff<sub>p<sub>Tet</sub></sub></b></td>
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<td class='row'><b>eff<sub>J23101</sub></b></td>
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<td class='row'><b>Declared efficiency</b></td>
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</tr><tr>
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<td class='row'>B0030</td><td class='row'>0.40</td><td class='row'>1.6814</td><td class='row'>2.45</td><td class='row'>0,6</td>
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</tr><tr>
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<td class='row'>B0031</td><td class='row'>0.01</td><td class='row'>ND</td><td class='row'>0.04</td><td class='row'>0,07</td>
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</tr><tr>
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<td class='row'>B0032</td><td class='row'>0.19</td><td class='row'>0.4193</td><td class='row'>0.40</td><td class='row'>0,3</td>
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</tr><tr>
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<td class='row'>B0034</td><td class='row'>1</td><td class='row'>1</td><td class='row'>1</td><td class='row'>1</td>
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</tr></table></div>
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<br><br>
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Estimated efficiencies in pSB4C5 plasmid with pTet-RBSx-GeneX-TT, with GeneX=mRFP, AiiA (<em> E. coli</em> TOP10, high copy number plasmid) or LuxI:
 +
<br><br>
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 +
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<div align="center"><table class='data' width='70%' border='1'><tr>
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<td class='row'><b>RBS</b></td>
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<td class='row'><b>eff<sub>mRFP</sub></b></td>
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<td class='row'><b>eff<sub>AiiA</sub></b></td>
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<td class='row'><b>eff<sub>LuxI</sub></b></td>
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<td class='row'><b>Declared efficiency</b></td>
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</tr><tr>
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<td class='row'>B0030</td><td class='row'>1.72</td><td class='row'>0.96</td><td class='row'>0.45</td><td class='row'>0,6</td>
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</tr><tr>
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<td class='row'>B0031</td><td class='row'>0.03</td><td class='row'>0.78</td><td class='row'>0.028</td><td class='row'>0,07</td>
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</tr><tr>
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<td class='row'>B0032</td><td class='row'>0.37</td><td class='row'>0.51</td><td class='row'>N.D.</td><td class='row'>0.3</td>
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</tr><tr>
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<td class='row'>B0034</td><td class='row'>1</td><td class='row'>1</td><td class='row'>1</td><td class='row'>1</td>
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</tr></table></div>
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</html>
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|}
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===Improvement===
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WHU-iGEM 2017
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https://parts.igem.org/Part:BBa_K2462006
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We improved this RBS by adding a promoter function in Bacillus mageterium.
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 +
 +
 
<!-- DON'T DELETE --><partinfo>BBa_B0030 EndReviews</partinfo>
 
<!-- DON'T DELETE --><partinfo>BBa_B0030 EndReviews</partinfo>

Latest revision as of 15:40, 27 October 2017

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Characterizations of BBa_B0030

The Alverno_Ca team characterized BBa_K2066046, which includes BBa_B0030, as well as a series of other plasmids using TX-TL, a mean of in vitro transcription and translation using cell extract. Below is a graph of normalized expression of BBa_K2066046 in comparison to the other palsmids in the sequence. T--Alverno_CA--W%26M_corrected.png

This graph shows the fluorescence of the plasmids, over a period of 12 and a half hours, in a plate reader. T--Alverno_CA--W%26M_plasmid_time_traces_in_TXTL%282%29.png"

Applications of BBa_B0030

William and Mary iGEM 2016

T--William_and_Mary--RBS_Characterization_Curves.png

We characterized the strengths of the Anderson RBS library.


User Reviews

UNIQ0b4225a0531c2896-partinfo-00000000-QINU

•••••

Antiquity

This review comes from the old result system and indicates that this part worked in some test.

•••

Aberdeen_Scotland 2009

The miniprep, single and double digests all worked. However we did not use this part for further cloning.

UNIQ0b4225a0531c2896-partinfo-00000003-QINU


UNIQ0b4225a0531c2896-partinfo-00000004-QINU

•••••

UNIPV-Pavia iGEM 2011

NB: unless differently specified, all tests were performed in E. coli MGZ1 in M9 supplemented medium at 37°C in low copy plasmid pSB4C5.

See Experience page of BBa_B0034 for more details.

Estimated efficiencies in pSB4C5 plasmid with -RBSx-mRFP-TT coding sequence under the control of the specified promoter:

RBS effpLux effpTet effJ23101 Declared efficiency
B00300.401.68142.450,6
B00310.01ND0.040,07
B00320.190.41930.400,3
B00341111


Estimated efficiencies in pSB4C5 plasmid with pTet-RBSx-GeneX-TT, with GeneX=mRFP, AiiA ( E. coli TOP10, high copy number plasmid) or LuxI:

RBS effmRFP effAiiA effLuxI Declared efficiency
B00301.720.960.450,6
B00310.030.780.0280,07
B00320.370.51N.D.0.3
B00341111

Improvement

WHU-iGEM 2017

https://parts.igem.org/Part:BBa_K2462006

We improved this RBS by adding a promoter function in Bacillus mageterium.


UNIQ0b4225a0531c2896-partinfo-00000007-QINU