Difference between revisions of "Team:Lethbridge/Measurement"

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                     <p>Our team decided to take part in this year’s interlab measurement study, measuring fluorescence of GFP in devices 1, 2 and 3. The GFP sequence was cleaved from I13504, using restriction enzymes Xba1 and Pst1, and ligated into pSB1C3 plasmid containing the following parts J23101, J23106 and J23117. Sequencing and restriction mapping was used to confirm proper insertion of the GFP sequence. Devices were transformed into DH5α cells. DH5α cells containing the devices and controls were normalized, lysed and GFP measurements were taken using a fluorometer.</p>
 
                     <p>Our team decided to take part in this year’s interlab measurement study, measuring fluorescence of GFP in devices 1, 2 and 3. The GFP sequence was cleaved from I13504, using restriction enzymes Xba1 and Pst1, and ligated into pSB1C3 plasmid containing the following parts J23101, J23106 and J23117. Sequencing and restriction mapping was used to confirm proper insertion of the GFP sequence. Devices were transformed into DH5α cells. DH5α cells containing the devices and controls were normalized, lysed and GFP measurements were taken using a fluorometer.</p>
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                    <p>Figure 1. Cell lysate fluorescence of iGEM Interlab study devices measured using fluorescence spectrophotometry (n = 3, mean ą s.d.)</p>
 
  
 
                     <p>The measurement of iGEM Interlab study devices yielded some unexpected results. Of the three devices, only Device 2 produced measurably different fluorescence levels than both the empty plasmid and no plasmid negative controls (Fig. 1). This questionable fluorescence data could be attributed to incorrect assembly of devices from the iGEM kit plates, as successful construct assembly was screened using restriction mapping only. In the future, it would be beneficial for iGEM headquarters to provide completed DNA constructs for large collaborative studies.</p>
 
                     <p>The measurement of iGEM Interlab study devices yielded some unexpected results. Of the three devices, only Device 2 produced measurably different fluorescence levels than both the empty plasmid and no plasmid negative controls (Fig. 1). This questionable fluorescence data could be attributed to incorrect assembly of devices from the iGEM kit plates, as successful construct assembly was screened using restriction mapping only. In the future, it would be beneficial for iGEM headquarters to provide completed DNA constructs for large collaborative studies.</p>
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<img src="https://static.igem.org/mediawiki/2015/9/9c/Uleth15_InterlabData1.png" height="40%" width="40%">
 
<img src="https://static.igem.org/mediawiki/2015/9/9c/Uleth15_InterlabData1.png" height="40%" width="40%">
 
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<p>Figure 1. Cell lysate fluorescence of iGEM Interlab study devices measured using fluorescence spectrophotometry (n = 3, mean ą s.d.)</p>
 
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Latest revision as of 00:13, 19 September 2015

iGEM

Measurement

Our team decided to take part in this year’s interlab measurement study, measuring fluorescence of GFP in devices 1, 2 and 3. The GFP sequence was cleaved from I13504, using restriction enzymes Xba1 and Pst1, and ligated into pSB1C3 plasmid containing the following parts J23101, J23106 and J23117. Sequencing and restriction mapping was used to confirm proper insertion of the GFP sequence. Devices were transformed into DH5α cells. DH5α cells containing the devices and controls were normalized, lysed and GFP measurements were taken using a fluorometer.

The measurement of iGEM Interlab study devices yielded some unexpected results. Of the three devices, only Device 2 produced measurably different fluorescence levels than both the empty plasmid and no plasmid negative controls (Fig. 1). This questionable fluorescence data could be attributed to incorrect assembly of devices from the iGEM kit plates, as successful construct assembly was screened using restriction mapping only. In the future, it would be beneficial for iGEM headquarters to provide completed DNA constructs for large collaborative studies.

Figure 1. Cell lysate fluorescence of iGEM Interlab study devices measured using fluorescence spectrophotometry (n = 3, mean ą s.d.)