-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.