Difference between revisions of "Part:BBa K1679006"
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===Overview===
 
===Overview===
 +
<p>This part can be added a promoter to meet a variety of requirements.</p>
 
<p>Promoter parts are often defined by a relatively short (~50 bp) sequence, but regions 100 bp or more upstream can affect promoter strength, and the effect of remote sequences can be reduced by including an insulator region.Ribo J is a kind of ribozyme-based insulator part that can buffer synthetic circuits from genetic context. It is a useful tool for measuring the strength of promoters which can make the disruption from genetic context to be reduced.
 
<p>Promoter parts are often defined by a relatively short (~50 bp) sequence, but regions 100 bp or more upstream can affect promoter strength, and the effect of remote sequences can be reduced by including an insulator region.Ribo J is a kind of ribozyme-based insulator part that can buffer synthetic circuits from genetic context. It is a useful tool for measuring the strength of promoters which can make the disruption from genetic context to be reduced.
 
As a matter of fact, our results from plate reader and flow cytometer show that the expression of GFP under J23117 promoter have a little difference with or without Ribo J. It proved that the Ribo J did has the function of buffering.</P>
 
As a matter of fact, our results from plate reader and flow cytometer show that the expression of GFP under J23117 promoter have a little difference with or without Ribo J. It proved that the Ribo J did has the function of buffering.</P>
 +
 +
===Plate Reader Result===
 +
 +
<h3>Equipment</h3>
 +
                <p>
 +
                    Varioskan Flash Multimode Reader<BR>Thermo Scientific<BR>Read Speed: 6.4 well per second
 +
                </p>
 +
                <h3>Software</h3>
 +
                <p>
 +
                    Run Software Version: Skanlt Software 2.4.5 RE for Varioskan Flash<BR>Current Software Version: Skanlt Software 2.4.5 RE for Varioskan Flash
 +
                </p>
 +
                <h3>Protocol</h3>
 +
                <p>
 +
                    Date: 2015 Aug 12<BR>1. Set our instrument to read OD600<BR>2. Setup a 96-well plate with our cultures<BR>3. Take the measurement and record it<BR>4. Calculate the dilution required for each sample (OD600=0.5)<BR>5. Dilute each sample<BR>6. Remeasure our sample on OD600<BR>7. Recalculate our dilution and remeasure until it's within 5% of 0.5.
 +
                </p>
 +
                <h3>Unit</h3>
 +
                <p>
 +
                    Three technical replicates of M9 liquid media was measured, which were called background value.<BR>A series of concentration of sodium fluorescein was measured, which are 0, 10, 20, 40, 80, 160, 320, 640 ng/mL . A calibration curve was defined.
 +
                </p>
 +
                <div style="width: 50%;margin: 0 auto;">
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                    <center>https://static.igem.org/mediawiki/2015/f/f5/OUC-China-InterLab_2.jpg</center>
 +
                </div>
 +
                <h3>Dataset</h3>
 +
                <p>
 +
                    All samples were cut the mean of background value, and then compared with the calibration curve to get the final dataset in units of fluorescein.
 +
                </p>
 +
               
 +
                <p></p>
 +
                <div style="width: 80%;margin: 0 auto;">
 +
                    <center>https://static.igem.org/mediawiki/2015/c/c5/OUC-China-InterLab_4.jpg</center>
 +
                </div>
 +
                <p></p>
 +
                <p>TR: Technical Replicate<BR>BR: Biological Replicate</p>
 +
                <div style="width: 100%;margin: 0 auto;">
 +
                    <center>https://static.igem.org/mediawiki/2015/4/4e/OUC-China-InterLab_5.png</center>
 +
                </div>
 +
                <p></p>
 +
  
 
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Revision as of 02:43, 19 September 2015

RiboJ and GFP

This part is a composite of RiboJ and I13504 which can be used to express GFP if added a promoter.

Overview

This part can be added a promoter to meet a variety of requirements.

Promoter parts are often defined by a relatively short (~50 bp) sequence, but regions 100 bp or more upstream can affect promoter strength, and the effect of remote sequences can be reduced by including an insulator region.Ribo J is a kind of ribozyme-based insulator part that can buffer synthetic circuits from genetic context. It is a useful tool for measuring the strength of promoters which can make the disruption from genetic context to be reduced. As a matter of fact, our results from plate reader and flow cytometer show that the expression of GFP under J23117 promoter have a little difference with or without Ribo J. It proved that the Ribo J did has the function of buffering.

Plate Reader Result

Equipment

Varioskan Flash Multimode Reader
Thermo Scientific
Read Speed: 6.4 well per second

Software

Run Software Version: Skanlt Software 2.4.5 RE for Varioskan Flash
Current Software Version: Skanlt Software 2.4.5 RE for Varioskan Flash

Protocol

Date: 2015 Aug 12
1. Set our instrument to read OD600
2. Setup a 96-well plate with our cultures
3. Take the measurement and record it
4. Calculate the dilution required for each sample (OD600=0.5)
5. Dilute each sample
6. Remeasure our sample on OD600
7. Recalculate our dilution and remeasure until it's within 5% of 0.5.

Unit

Three technical replicates of M9 liquid media was measured, which were called background value.
A series of concentration of sodium fluorescein was measured, which are 0, 10, 20, 40, 80, 160, 320, 640 ng/mL . A calibration curve was defined.

OUC-China-InterLab_2.jpg

Dataset

All samples were cut the mean of background value, and then compared with the calibration curve to get the final dataset in units of fluorescein.

OUC-China-InterLab_4.jpg

TR: Technical Replicate
BR: Biological Replicate

OUC-China-InterLab_5.png


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 745