Difference between revisions of "Team:NTU SINGAPORE/Collaborations"

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<p>Finally, use ΔCt(test)2- ΔCt(control)1 to get ΔΔCt.</p>
 
<p>Finally, use ΔCt(test)2- ΔCt(control)1 to get ΔΔCt.</p>
  
<p>As shown from the above bar charts, 20mM of IPTG induction has greater fold change of gene expression than 200mM of IPTG induction. It is worth to notice that the RT-PCR efficiency might be different for different genes due to their size of PCR products and the formula used above is assuming the PCR efficiency to be 100% that eventually resulted in 2 copies of products.</p>
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<p>5mM of IPTG and 20mM of glucose was used to induce the gene expression. It is worth to notice that the RT-PCR efficiency might be different for different genes due to their size of PCR products and the formula used above is assuming the PCR efficiency to be 100% that eventually resulted in 2 copies of products.</p>
  
  

Revision as of 09:14, 23 October 2017



Macquaire iGEM Team


Macquaire team had constructed a hydrogenase gene capable of converting glucose to hydrogen gas. The plasmid (BBa_KXXXXXX) that consist of hydrogenase enzyme (Hyd1), ferredoxin, ferredoxin-NADPH-reductase (FNR) and maturation enzyme (HydEF and HydG), in which their expression are co-regulated under pLac promoter.


This year again, our team helped them to evaluate each of the gene expression level by using RT-PCR.


The results of RT-PCR is shown as below:

Formula: 2^-(ΔΔCt) is used to calculate the fold change of gene expression after induction, in which ΔCt(control)1= Ct(uninduced operon genes)- Ct(Chloramphenicol gene)ΔCt(test)2 = Ct(Induced operon genes)-Ct(Induced Chloramphenicol gene)


Finally, use ΔCt(test)2- ΔCt(control)1 to get ΔΔCt.

5mM of IPTG and 20mM of glucose was used to induce the gene expression. It is worth to notice that the RT-PCR efficiency might be different for different genes due to their size of PCR products and the formula used above is assuming the PCR efficiency to be 100% that eventually resulted in 2 copies of products.