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Team:Tokyo Metropolitan/Notebook/A22
From 2011.igem.org
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Treated restriction enzyme
Electrophoresis
to oriTr-GFP connected parts refined on 17 Aug.
Details
<Treated restriction enzyme>
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.
