Difference between revisions of "Team:Gifu/result-page"

PROJECT

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RESULT

EFFICIENCY

Summary of the experiment

  The existence of circular mRNA is confirmed by RNase processing. Endogenous RNA (linear RNA)(GAPDH) is decomposed by RNase R (exoribonuclease), but circular RNA is not decomposed.
  Double-stranded DNA from undecomposed RNA can be gained with RT-PCR. So the existence of circular mRNA is confirmed by the observation of the DNA with electrophoresis.

Flow of the experiment

Purpose: proving the existence of circular mRNA
Goal: finding the RNA that is decomposed by endoribonuclease but is not decomposed by exoribonuclease
Protocol:
  1. RNase processing: to find the circular mRNA
  2. RT-PCR: to synthesize cDNA and to detect the cDNA
  3. synthesized from circular mRNA or endogenous RNA
Electrophoresis: to detect the DNA synthesized from the cDNA

   Ct is a value that the more the gene template in PCR increases, the more Ct value decreases. If there is a difference between “C” and “D”, there is a difference in the quantity of template geneThus, the difference between “C” and “D” is small; the cyclization may be efficiency.As a result, Ct in 2014 was 2.31, but “outside”, “inside①”, “inside②” were 1.28, 2.02, 1.89 collectively. From these things, it can be said that the efficiency of cyclization rose with all devices which we designed in this time. Especially, in the case of “outside”, there was a difference with Ct more than 1 point than “2014”. When we think that a quantity of the gene doubles for 1 cycle in PCR simply, it can be said that the cyclic efficiency of “outside” was twice as high as that of “2014”.
  After all, it is thought that it fitted this splice site because it is a complementarity chain derived from the creature.

FUNCTION