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Cloning Vector Map

Cloning Vector Map. Vector Characteristics. Topoisomerase. Contains a T-overhang Linked topoisomerase. Topoisomerase. PCR Fragment Properties. A. PCR Product. A. Contains an A-overhang A-T base pairing helps anchor the insert to vector. Topoisomerase. PCR Product. A. A.

JasminFlorian
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Cloning Vector Map

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  1. Cloning Vector Map

  2. Vector Characteristics Topoisomerase • Contains a T-overhang • Linked topoisomerase Topoisomerase

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

  4. Topoisomerase PCR Product A A Topoisomerase Ligation Mix • 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)

  5. How it Works?

  6. Recombinant Construct

  7. 0 Transformation Competent Cells (E. coli Heat Shock Method

  8. 0 Cloning

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