Revision as of 13:00, 11 April 2012

PcTF Subcloning in E. coli

11/4/12

> Digests (Fermentas FD)
BD-112 and BD-113

White dashed lines border show the vector backbone and the BD-111 constructed gene(RBS + PcTF).

Plasmid DNA	2.0 μl*
EcoR1	1.0 μl
Pst1	1.0 μl
10x FastDigest buffer + green loading dye	1.5 μl
dH₂O	9.5 μl
	15.0 μl total
Incubate at 37°C for 10 minutes.

Make the (1%) agarose glee and add 15μl of restricted DNA (PcTF & RBS) in one well and 10 μl of ladder.

'The size of the constructed gene supposed to be: 1123 + 15 + 6 = 1144 bp'

@@ Line 7: / Line 7: @@
 <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
-=Date=
+=11/4/12=
-'''List title'''
+> Digests (Fermentas FD)<br>
-# List items
+BD-112 and BD-113
+[[Image:BD-111-Behzad-5-4-12.jpg|thumb|350px|White dashed lines border show the vector backbone and the BD-111 constructed gene(RBS + PcTF).]]
+{| class="wikitable" width=400px
+| Plasmid DNA || 2.0 μl*
+|-
+| EcoR1  || 1.0 μl
+|-
+| Pst1 || 1.0 μl
+|-
+| 10x FastDigest buffer + green loading dye || 1.5 μl
+|-
+| dH<sub>2</sub>O || 9.5 μl
+|-
+| &nbsp; || 15.0 μl total
+|-
+| Incubate at 37°C for 10 minutes.
+|}
+# Make the (1%) agarose glee and add 15μl  of restricted DNA (PcTF & RBS) in one well and 10 μl  of ladder.
+'The size of the constructed gene supposed to be: 1123 + 15 + 6 = 1144 bp'