Team:Tokyo Metropolitan/Notebook/A22

From 2011.igem.org

(Difference between revisions)
(-Targeting-)
(-Visualization-)
 
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= -Visualization- =
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Treated restriction enzyme
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Digestion
Electrophoresis
Electrophoresis
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Details
Details
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<Treated restriction enzyme>
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<Digestion>
Refered to DNA concentration mesured on 17 Aug and bands checked on 18 Aug, mixed reagents as follows. And EcoR I or pst I might were inactivation, thus made only EcoR I and pst I.
Refered to DNA concentration mesured on 17 Aug and bands checked on 18 Aug, mixed reagents as follows. And EcoR I or pst I might were inactivation, thus made only EcoR I and pst I.
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Checked a band same position (3 kb) on 18 Aug for treated with only EcoR I and only pst I, two bands (2.2 kb and 1.3 kb) for treated with EcoR I + pst I.  
Checked a band same position (3 kb) on 18 Aug for treated with only EcoR I and only pst I, two bands (2.2 kb and 1.3 kb) for treated with EcoR I + pst I.  
 +
Therefore it was successful to ligate for oriTr-GFP. And the restriction enzyme was not inactivation.
Therefore it was successful to ligate for oriTr-GFP. And the restriction enzyme was not inactivation.

Latest revision as of 10:37, 5 October 2011

-Visualization-

Digestion

Electrophoresis

to oriTr-GFP connected parts refined on 17 Aug.


Details

<Digestion>

Refered to DNA concentration mesured on 17 Aug and bands checked on 18 Aug, mixed reagents as follows. And EcoR I or pst I might were inactivation, thus made only EcoR I and pst I.


oriTr-GFP1-E

 EcoR 1µl

 plasmid 4µl

 DW 10.75µl

 10×H 1.75µl

total 17.5µl


oriTr-GFP1-P

 Pst I 1µl

 plasmid 4µl

 DW 10.75µl

 10×H 1.75µl

total 12.5µl


oriTr-GFP1-EP

 EcoR I 1µl

 PstⅠ 1µl

 plasmid 4µl

 DW 9.75µl

 10×H 1.75µl

total 17.5µl


Also made same three types solution for oriTr-GFP 2.


<Electrophoresis>

According to protocol.(cf. lab notebook on 3 Aug)

Marker was 5 µl : Gene Ladder Wide 1 (borrowed from Plant hormone lab)


Results

This time made good results.

Checked a band same position (3 kb) on 18 Aug for treated with only EcoR I and only pst I, two bands (2.2 kb and 1.3 kb) for treated with EcoR I + pst I.

Therefore it was successful to ligate for oriTr-GFP. And the restriction enzyme was not inactivation.