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