Cloning Vector Map.

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Presentation transcript:

Cloning Vector Map

Vector Characteristics Topoisomerase Contains a T-overhang Linked topoisomerase Topoisomerase

PCR Fragment Properties PCR Product A Contains an A-overhang A-T base pairing helps anchor the insert to vector

Ligation Mix Single 3´ thymidine (T) overhangs for TA Cloning Topoisomerase PCR Product A Single 3´ thymidine (T) overhangs for TA Cloning Taq DNA polymerase adds nontemplate dependent A at the 3’ end Topoisomerase covalently bound to the vector ("activated“ vector)

How it Works?

Recombinant Construct

Transformation Competent Cells (E. coli Heat Shock Method

Cloning

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