Team:UESTC-China/Protocol

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UESTC-China

DNA ligation with T4 DNA Ligase
Component 20ul Reaction
1.33X Master Mix 15ul
Insert DNA the moles of insert DNA to vector DNA is 5:1
>Vector DNA 50ng
ddH2O To 20ul
Incubate at 50℃ for 1 hour
DNA ligation with Gibson Assemble
Component 20ul Reaction
Vector DNA 50ng
Insert DNA the moles of insert DNA to vector DNA is 5:1
10X T4 DNA ligase buffer 2ul
T4 DNA ligase 1ul
ddH2O to 20ul
Incubate at 25℃ for 12 hours