Toggle navigation SCU-Igem Project Background Overview Description Result Modeling Human Practice Presentation at Seventh Senior High School Visiting University of Electronic Science and Technology of China (UESTC) Safety Parts Attributions Team Notebook Notebook Notebook of Biobricks Notebook of Transmitter Notebook of Effector Notebook of Sensor Method Bacterial Genomic DNA Prep Digestion Gel Extraction Linkage PCR Plasmid Mini Prep Linkage Protocol Back to top According to the advice of IGEM Competition, there are two linkage methods that we use in experiment. The first one is “3A” Assembly and the second is “Traditional” Assembly. For “3A” Assembly, we set up the following system (20 μl):Linear Backbones from IGEM Competition( with K+, C+, A+, or T+) About 100 ng Insert DNA 1 The molar ratio of Insert DNA to Backbone is 3:1 to 5:1 Insert DNA 2 Same to Insert DNA 1 10 x T4 DNA Ligase Buffer 2 μl T4 DNA Ligase 1 μl H2O Up to 20 μl For “Traditional” Assembly, we set up the following system (20 μl):Linear Backbones from IGEM Competition( with K+, C+, A+, or T+) About 100 ng Insert DNA The molar ratio of Insert DNA to Backbone is 3:1 to 5:1 10 x T4 DNA Ligase Buffer 2 μl T4 DNA Ligase 1 μl H2O Up to 20 μl Sichuan university
Linkage Protocol
According to the advice of IGEM Competition, there are two linkage methods that we use in experiment. The first one is “3A” Assembly and the second is “Traditional” Assembly.
Linear Backbones from IGEM Competition( with K+, C+, A+, or T+)
About 100 ng
Insert DNA 1
The molar ratio of Insert DNA to Backbone is 3:1 to 5:1
Insert DNA 2
Same to Insert DNA 1
10 x T4 DNA Ligase Buffer
2 μl
T4 DNA Ligase
1 μl
H2O
Up to 20 μl
Insert DNA
Sichuan university
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