IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Output/2009/06/30/Zhangs

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Double digestion of CI
System for front insert:

10μL plasmid
2μL 10×H beffer
1.5μL EcoRI
1.5μL SpeI
5μL ddH2O
20μL Total

4 h digested
Electrophoresis
Result (from left to right)
CI, CI plasmid control, marker
PKU 20090630 Zhangs 1.JPG
Gel: 0.072gPN volume: 216μL
Gel extraction for CI insert

Ligation Insert:CI Vector:1-23L, 1-4H
System
(1)7:1 system

7μL CI insert
1μL vector
1μL 10× Ligase buffer
1μL Ligase
10μL Total

(2)3:1 system

3μL CI insert
1μL vector
1μL 10× Ligase buffer
1μL Ligase
4μL ddH2O
10μL Total

Control

0 CI insert
1μL vector
1μL 10× Ligase buffer
1μL Ligase
7μL ddH2O
10μL Total

6 samples in total, 2 for 1-23L, 2 for 1-4H, 2 for 1-23L & 1-4H controls