Team:Tokyo Metropolitan/Notebook/S27
From 2011.igem.org
-Targeting-
-Plasmid extraction
Extracted plasmids of Anti-killer Device(6)(pSB1C3), Anti-killer Device(7)(pSB1C3), J23100-P0140(J61002), AHL induce Device(pSB1C3) and I13522.
-Digestion
Digested J23100-P0140(J61002), AHL indece Device(pSB1C3) and I13522.
Recipes
J23100-P0140(J61002)
DNA 20ul
DW 21ul
Buffer 5ul
XbaI 2ul
PstI 2ul
AHL induce Device(pSB1C3)
DNA 10ul
DW 31ul
Buffer(H) 5ul
SpeI 2ul
PstI 2ul
I13522
DNA 20ul
DW 21ul
Buffer(M) 5ul
XbaI 2ul
PstI 2ul
-Electrophoresis
Electrophoresed J23100-P0140(J61002), AHL induce Device(pSB1C3) and I13522.
Results
Did not show all band.
-Pre-culture
Pre-cultured Anti-killer Device⑥(pSB1C3), Anti-killer Device⑦(pSB1C3), J23100-P0140(J61002), AHL induce Device(pSB1C3) and I13522.
-Visualize-
-oriTr-GFP, oriTr electrophoresis
-oriTr-GFP, oriTr Gel extraction
-oriTr-GFP, oriTr Measurement of DNA concentration
-oriTr-GFP + oriTr Ligation
-oriTr-GFP + oriTr Transformation
-Speeding up-
Colony PCR of BBa_K543002
Electrophoresis