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==1.3. DNA hybridization in solution of H<sub>2</sub>O<sub>2</sub>==
:&nbsp;&nbsp;&nbsp;&nbsp;H<sub>2</sub>O<sub>2</sub> has strong corrosion. There was a risk that DNA strand is denatured by H<sub>2</sub>O<sub>2</sub><sup>[1]</sup>. So, we had to ensure that the hybridization of DNA does not suffer from H<sub>2</sub>O<sub>2</sub>. We guessed that if DNA had been destroyed, then molecular mass would decrease in that DNA can't hybridize. Visualizing the relative magnitude of the molecular weight by electrophoresis, we are able to examine whether DNA can hybridize or not.<br>
:&nbsp;&nbsp;&nbsp;&nbsp;This time, we have to examine the state of the DNA in the time we need to steer rocket, set the time to soak and  concentration of H<sub>2</sub>O<sub>2</sub>. We used PAGE electrophoresis to ascertain the stability of DNA duplex in thin H₂O₂ solution 1%-5%. PAGE electrophoresis shows the difference of molecular weight that comes from dissociation or hybridization in the form of bands.<br><br>
:Reference<br>
:[1]Natacha Driessens, Soetkin Versteyhe, Chiraz Ghaddhab, Agne`s Burniat, Xavier De Deken, Jacqueline Van Sande, Jacques-Emile Dumont, Franc¸oise Miot, Bernard Corvilain,Endocrine-Related Cancer 16 845–856(2009)<br>


=The result of DNA hybridization in solution of hydrogen peroxide=
::::::::::>>back to [[Biomod/2012/Titech/Nano-Jugglers/Results#1.3._DNA_hybridization_in_solution_of_H2O2|Results]]
:'''>>Result page is [[Biomod/2012/Titech/Nano-Jugglers/Results/DNA_hybridization|here]]'''
 
 
:Image 1 and 2 shows the observation results of DNA hybridization in solution of Hydrogen peroxide for 90minutes.
:From the pictures, the hybridized DNA band appear in lane 1,2,3&8. In lane 4,5,6&7 appears single strand band.
:Lane1&8 are the same, lane 2 & 3 are added hydrogen peroxide solution (1% and 5% for 1h)for 90 minutes. In Lane 5&7 shows the observation results of single strand DNA (ssDNA) in solution of Hydrogen peroxide. In lane 4&6 are the control band of ssDNA.
 
 
:White bar in Image2 represents the same position of Lane 1&8 in that they are the same sample.
:Comparing between 4.5.6&7, these 4 lines appear lower than the white line, and 4-5and 6-7 are few differences. If hydrogen peroxide affects ssDNA, destroy, tear up or denature, line 5and 7 will appear in the lower position or becomes unclear. Judging from appearances,  differences between positive and negative control ware few.
:Comparing 1.2.3 and 8,these lines are completely appear in the white bar.
:Differences between these lines are only concentration of hydrogen peroxide. So, we conclude that there is no influence of hydrogen peroxide for DNA hybridizations however DNA is exposed to hydrogen peroxide within 90minutes.
:So, we could determine that there is no effect of hydrogen peroxide for dsDNA as well as well as ssDNA.
 
R: SAWADAY ND14 (25mer) DNA
CAAAAGAGTTAGGATGGGAGCTGGA
 
D: SAWADAY D14(25mer) DNA
TCCAGGTCCCATCCTAACTCTTTTG
Hydrogen peroxide concentration(w/v)
 
①D+R, 0% Hydrogen peroxide
②D+R, 1% Hydrogen peroxide
③D+R, 5% Hydrogen peroxide
④D, 0% Hydrogen peroxide
⑤D, 5% Hydrogen peroxide
⑥R, 0% Hydrogen peroxide
⑦R, 5% Hydrogen peroxide
⑧D+R, 0% Hydrogen peroxide
⑨

Latest revision as of 23:16, 27 October 2012


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1.3. DNA hybridization in solution of H2O2

    H2O2 has strong corrosion. There was a risk that DNA strand is denatured by H2O2[1]. So, we had to ensure that the hybridization of DNA does not suffer from H2O2. We guessed that if DNA had been destroyed, then molecular mass would decrease in that DNA can't hybridize. Visualizing the relative magnitude of the molecular weight by electrophoresis, we are able to examine whether DNA can hybridize or not.
    This time, we have to examine the state of the DNA in the time we need to steer rocket, set the time to soak and concentration of H2O2. We used PAGE electrophoresis to ascertain the stability of DNA duplex in thin H₂O₂ solution 1%-5%. PAGE electrophoresis shows the difference of molecular weight that comes from dissociation or hybridization in the form of bands.

Reference
[1]Natacha Driessens, Soetkin Versteyhe, Chiraz Ghaddhab, Agne`s Burniat, Xavier De Deken, Jacqueline Van Sande, Jacques-Emile Dumont, Franc¸oise Miot, Bernard Corvilain,Endocrine-Related Cancer 16 845–856(2009)
>>back to Results
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