Revision as of 16:57, 12 January 2013

PcTF Subcloning in E. coli

01/12/2013

Ligation of KAH111, KAH112 and C-His vector in DH5α-T and BL21

The 4 μL of C-His Vector from stock is diluted with 20 μL of nuclease free water, then cut with E/P and purified from the gel.
Ligations

Ligation	Plate results (lig : neg crtl) 12/12/12
1. E. coli DH5α-T KAH111(E/P)/size, 20 ng + C-His(E/P)/3600, 26ng	KAH111/C-His 3:1 No Colonies
2. E. coli BL-21 KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng	KAH111/C-His 3:1 No Colonies
3. C-His(E/P)/ 25 ng

@@ Line 7: / Line 7: @@
 <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
-=Date=
+=01/12/2013=
+Ligation of KAH111, KAH112 and C-His vector in DH5α-T and BL21
+* The 4 μL of C-His Vector from stock is diluted with 20 μL of nuclease free water, then cut with E/P and purified from the gel.
+* Ligations
+{| {{table}} cellspacing="3" <!-- Ligations table -->
+|- bgcolor=#cfcfcf
+| Ligation || <font color="blue"><u>Plate results (lig : neg crtl)</u> 12/12/12</font>
+|-
+| 1. E. coli DH5α-T KAH111(E/P)/size, 20 ng + C-His(E/P)/3600, 26ng || <font color="blue">KAH111/C-His 3:1 No Colonies</font>
+|-
+| 2. E. coli BL-21   KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng || <font color="blue">KAH111/C-His 3:1 No Colonies</font>
+|-
+| 3. C-His(E/P)/ 25 ng ||
+|}
-'''List title'''
+{| {{table}} cellspacing="3" <!-- Ligation rxn table -->
-# List items
+| &nbsp;             || 1    || 2  || 3
+|-
+| Insert DNA         || 3.12  || 3.12 || ---
+|-
+| Vector DNA         || 2.94  || 2.94 || 2.94
+|-
+| 2x lgn buf (Roche) || 7.06  || 7.06 || 5.0
+|-
+| T4 ligase (NEB)    || 1.0  || 1.0  || 1.0
+|-
+| dH<sub>2</sub>O    || 0 || 0 || 0.88
+|-
+| &nbsp;             || 14.12 μL || 14.12 μL || 10μl
+|}