Revision as of 14:06, 12 September 2008

Checking our ligases (following)

Ligation of the plasmid digested by EcoRI

Ligation of the plasmid digested by BspHI

1% agarose gel

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	MP101	MP101	D202	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	MP101	MP101	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

@@ Line 65: / Line 65: @@
 |colspan="6"|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)
+{|border="1" style="text-align: center"
+|'''well n°'''
+|1
+|2
+|3
+|4
+|5
+|6
+|7
+|8
+|9
+|10
+|11
+|-
+|'''sample'''
+|rowspan="3"|1 kb ladder
+|rowspan="3"|MP101
+|rowspan="3"|MP101
+|rowspan="3"|D203
+|'''8'''
+|'''9'''
+|'''10'''
+|'''11'''
+|'''12'''
+|'''13'''
+|rowspan="3"|100 bp ladder
+|-
+|'''purification <br> digestion products'''
+|colspan="3"|column
+|colspan="3"|no <br> purification
+|-
+|'''ligase used'''
+|L1
+|L2
+|L3
+|L1
+|L2
+|L3
+|-
+|'''amount loaded'''
+|10 µL
+|0,3 µL
+|0,5 µL
+|4 µL
+|colspan="3"|15 µL
+|colspan="3"|10 µL
 |10 µL
 |}