Latest revision as of 14:16, 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 of 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 of 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 7: / Line 7: @@
 % agarose gel
-[[Image:Ligation-D202.JPG|thumb|Ligation of the plasmid digested by EcoRI]]
+[[Image:Ligation-D202.JPG|thumb|Ligation of the plasmid digested by '''EcoRI''']]
-[[Image:Ligation-D203.JPG|thumb|Ligation of the plasmid digested by BspHI]]
+[[Image:Ligation-D203.JPG|thumb|Ligation of the plasmid digested by '''BspHI''']]
 '''Analysis of the ligation of the plasmid digested by EcoRI'''
@@ Line 43: / Line 43: @@
 |rowspan="3"|100 bp ladder
 |-
-|'''purification <br> digestion products'''
+|'''purification of <br> digestion products'''
 |colspan="3"|gel extraction
 |colspan="3"|column
@@ Line 98: / Line 98: @@
 |rowspan="3"|100 bp ladder
 |-
-|'''purification <br> digestion products'''
+|'''purification of <br> digestion products'''
 |colspan="3"|column
 |colspan="3"|no <br> purification