Difference between revisions of "Team:Gifu/More/"

Inter Lab Study

Equipment

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• BioSHAKER TA-12R
  
use for shaking culture




• Safire ART-NR:F129013
  
use for measurement of absorbance

Method of Measurement

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  We constructed 5 plasmids [positive control, negative control, device1~3] by using BioBricks and then transformed to E. coli K-12 JM109. We checked them by colony direct PCR and plasmids sequence. We cultivated correct bacterias by shaking until OD660:0.5. Cultivated mediums were　transferred to 96-well plate by 100mL. We read the absorbance.


  Setting of plate reader is following:




Sequence data

・device 1 : J23101+ I13504


・device 2 : J23106+ I13504


・device 3 : J23117+ I13504




These sequence datas are correct.

Result

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• Definition
  

  We expressed absorbance as relative value for　positive control.
This expression is following:



  Sample : Absorbance in sample
  NCav : Average of absorbance in negative control
  PCav : Average of absorbance in positive control







• Value
  

Relative absorbance values for positive control are shown below.









FIG1 Relative absorbance values
20A : Device1 (J23101 + I13504)
22A : Device2 (J23106 + I13504)
22K : Device3 (J23117 + I13504)

Discussion

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  The value of device1 is much higher than that of device2 and 3. This is why each device is different from promoter sequence, especially -10 and -35 region.







Promoter sequence in device1 resembles consensus sequence. On the other hand, device2 and 3 is not similar than device1. This is how each GFP expression is different.