Biomod/2013/Titech/see more 9

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<a href="http://openwetware.org/wiki/Biomod">Biomod<a/>|<a href="http://openwetware.org/wiki/Biomod/2013">2013</a>


    RNaseH is ribonuclease that degrades RNA in DNA-RNA double strand and release single strand DNA. RNaseH is a nonspecific endo nuclease and degrades RNA by hydrolysis reaction. Mg2+, which is bivalent metalion connecting with enzyme, helps activation of RNaseH.


Unit definition
One unit is the amount of the enzyme that produces 1 nmol of acid-soluble 3H in 20 minutes at 30°C and pH7.7, with poly (rA)・poly (dT) as the substrate.


    We calculated following above definition and got the result that the reaction between RNaseH and that double strand finishes in twenty minutes when the concentration of DNA-RNA double strand is 1μL in the 5μL solution, and RNaseH is 0.1U. In the present experiment, we did an experiment using 0.5U RNaseH.

 

    RNaseH doesn’t degrade single strand RNA. We designed Linker complex as RNA portion is not degraded by RNaseH when Linker complex is inactivated. To realize that, we made RNA portion rope shape that can keep single strand.

<img src="https://upload.wikimedia.org/wikipedia/commons/2/26/See_more_method_24.png" width="330"> <img src="https://upload.wikimedia.org/wikipedia/commons/f/f0/111111%E3%81%86%E3%81%A4%EF%BD%99%EF%BD%87%EF%BD%86%EF%BD%8B%EF%BD%99%EF%BD%87%E3%81%8D%E3%81%86%EF%BD%87.png" width="330">

Fig. 1. Sequence of Linker-output complex

 

 


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