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STEPS IN GENE CLONING - Coggle Diagram
STEPS IN GENE CLONING
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CLEAVE OR CUT
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SINGLE CUT, WITHIN THE lacZ GENE, CAUSED THE PLASMIDS TO OPEN AND THE DISRUPTION OF THE GENE
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INSERTION
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MIX THE CUT PLASMID AND DNA FRAGMENTS ALLOWING BASE PAIR BETWEEN THEIR COMPLEMENTARY STICKY ENDS BY FORMING HYDROGEN BONDS.
DNA LIGASE IS ADDED TO SEAL THEM TOGETHER AT THE SUGAR PHOSPHATE BACKBONES OF THE FRAGMENTS WHOSE STICKY ENDS HAVE BASE-PAIRED WITH PHOSPHODIESTER BOND. THUS, FORMING A RECOMBINANT DNA.
ISOLATION
ISOLATION OF TARGET DNA OF INTEREST FROM SOURCES. Eg: HUMAN CELLS, PLANT CELLS AND ANIMAL CELLS.
TO ISOLATE A SPECIFIC GENE, ONE OFTEN BEGINS BY CONSTRUCTING A DNA LIBRARY- A COMPREHENSIVE COLLECTION OF CLONED DNA FRAGMENTS FORM A CELL, TISSUE OR ORGANISM.
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