Cloning Vector Map by 8657lW88

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									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
                  Ligation Mix

Topoisomerase
                       PCR Product   A
                   A




                              • Single 3´ thymidine (T)
  Topoisomerase
                                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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