IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/07/06/Zhangs

Identify the RBS+CI+T ligation product by electrophoresis
Result (from left to right)
Marker, RBS plasmid control, 1-5J+CI+T①~⑤ (plasmid), 1-2I+CI+T①~⑤ (plasmid), 1-11N+CI+T①~⑤ (plasmid)

The ligation of 1-5J+CI+T failed
Marker, RBS plasmid control, 1-1H+CI+T①~⑤ (plasmid), 1-2G+CI+T①~⑤ (plasmid)

The 1-1H+CI+T1, 5 & 1-2G+CI+T1, 2, 3, 5 are the target plasmids.
Marker, RBS plasmid control, 1-5N+CI+T①~⑤ (plasmid)

The 1-5N+CI+T2, 4, 5 are the target plasmids.

Choose samples:
4μL 1H+CI+T⑤, 4μL 2G+CI+T⑤, 6μL 5N+CI+T⑤, 6μL 2I+CI+T⑤, 6μL 11N+CI+T④
Concentration
217.5ng/μL 233ng/μL 125ng/μL 162ng/μL 123ng/μL

Double-digestion of CI+1-23L insert (causing the ligation of 1-5J+CI+T failed).

1.5μL	XbaI
1.5μL	PstI
2μL	10×M buffer
	Plasmid
	ddH2O
20μL	Total

4 hours digesting.
Electrophoresis to purify the CI+1-23L insert, and extraction.