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TCTAGA

TCTAGA. AGATCT. T CTAGA. 3’. 5’. AGATC T. 3’. 5’. DNA CLONING: RESTRICTION ENZYMES. 3’. 5’. XbaI. 5’. 3’. T. CTAGA. 5’. 3’. AGATC. T. 3’. 5’. T. CTAGA. 5’. 3’. T. AGATC. 3’. 5’. DNA CLONING: DNA LIGASE. -OH. P-. -P. OH-. 2 ATP. DNA ligase + ATP.

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TCTAGA

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  1. TCTAGA AGATCT T CTAGA 3’ 5’ AGATC T 3’ 5’ DNA CLONING: RESTRICTION ENZYMES 3’ 5’ XbaI 5’ 3’

  2. T CTAGA 5’ 3’ AGATC T 3’ 5’ T CTAGA 5’ 3’ T AGATC 3’ 5’ DNA CLONING: DNA LIGASE -OH P- -P OH- 2 ATP DNA ligase + ATP 2 AMP +2PPi

  3. DNA CLONING: plasmid vectors bacterial plasmid E. coli multiple cloning site - PstI - XbaI origin of replication (ori) gentamycin resistance gene (gent)

  4. DNA CLONING: TRANSFORMATION “VECTOR” + + “COMPETENT CELLS” Chemically treated to enhance DNA uptake E. coli “TRANSFORMED” BACTERIA

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