Revision as of 14:33, 4 October 2013

Digestion and Ligation

Digestion :


Enzyme	EcoRI	PstI	SpeI	XbaI
Buffer	Fast Digest	Fast Digest	Fast Digest	Fast Digest
Site	5'-G/AATTC-3' 3'-CTTAA/G-5'	5'CTGCA/G 3' 3'G/ACGTC 5'	5'A/CTAGT 3' 3'T G ATC/A5'	5'T/CTAGA3' 3'AGATC/T5'

Double Digestion :

Simple Digestion :

1 .Incubate at 37°C for 10-90min 2. Inactivate digestion of enzymes by ethanol precipitation

Ligation:

Mix in a molar ratio the insert and the linear vector

@@ Line 14: / Line 14: @@
 ! scope=col | XbaI
 |-
-| width="33%" |
+| width="30%" |
-Tampon
+Buffer
-| width="34%" |
+| width="30%" |
 Fast Digest
-| width="33%" |
+| width="30%" |
 Fast Digest
-| width="34%" |
+| width="30%" |
 Fast Digest
-| width="33%" |
+| width="30%" |
 Fast Digest
 |-
 | width="33%" |
-site
+Site
 | width="34%" |
 '-G/AATTC-3'
@@ Line 54: / Line 54: @@
 |-
 | width="33%" |
-Tampon FD 10X
+Buffer Fast Digest 10X
 | width="34%" |
 µL
@@ Line 87: / Line 87: @@
 |-
 | width="33%" |
-Tampon FD 10X
+Buffer Fast Digest 10X
 | width="34%" |
 µL
@@ Line 107: / Line 107: @@
-.Incubate at 37°C during 10-90min
+.Incubate at 37°C for 10-90min
-. Inactivate digestion enzyms with ethanol precipitation
+. Inactivate digestion of enzymes by ethanol precipitation
 Ligation:
 Mix in a molar ratio the insert and the linear vector