Revision as of 14:13, 12 September 2008

Checking our ligases (following)

1% agarose gel

Ligation of the plasmid digested by EcoRI

Ligation of the plasmid digested by BspHI

Analysis of the ligation of the plasmid digested by EcoRI
No heat inactivation of the ligase

well n°	1	2	3	4	5	6	7	8	9	10	11	12	13
sample	1 kb ladder	circular plasmid (MP101)	circular plasmid (MP101)	digested plasmid (D202)	digested plasmid (D202)	1	2	3	4	5	6	7	100 bp ladder
purification digestion products						gel extraction			column			no purification
ligase used						L1	L2	L3	L1	L2	L3	L1
amount loaded	10 µL	0,5 µL	1 µL	2 µL	3 µL	10 µL						3 µL	10 µL

Analysis of the ligation of the plasmid digested by BspHI
Heat inactivation of the ligase (20 min at 65°C)

well n°	1	2	3	4	5	6	7	8	9	10	11
sample	1 kb ladder	circular plasmid (MP101)	circular plasmid (MP101)	digested plasmid (D203)	8	9	10	11	12	13	100 bp ladder
purification digestion products					column			no purification
ligase used					L1	L2	L3	L1	L2	L3
amount loaded	10 µL	0,3 µL	0,5 µL	4 µL	15 µL			10 µL			10 µL

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+<br>
 '''Analysis of the ligation of the plasmid digested by BspHI'''
 <br>Heat inactivation of the ligase (20 min at 65°C)