I really need help I can't find it in my notes online and I need to know this for a test
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-Identify and Isolate the gene you want on an mRNA strand
-Remove that mRNA and place it in a solution containing free nucleotides, and the enzyme reverse transcriptase to produce cDNA
-Then mix the cDNA with a restriction endonuclease enzyme to create sticky ends.
-Then you prepare the vector. You take a plasmid (or whatever vector you choose to use) and mix it with the SAME restriction endonuclease enzyme to create complementary sticky ends to that of the gene.
-Then mix the gene and vector together in a solution containing the enzyme DNA ligase. This encourages hydrogen bonding between the complementary nucleotides of the gene and plasmid.
-You have now created recombinant DNA which can be inserted in to which ever organism you which to modify.
-Remove that mRNA and place it in a solution containing free nucleotides, and the enzyme reverse transcriptase to produce cDNA
-Then mix the cDNA with a restriction endonuclease enzyme to create sticky ends.
-Then you prepare the vector. You take a plasmid (or whatever vector you choose to use) and mix it with the SAME restriction endonuclease enzyme to create complementary sticky ends to that of the gene.
-Then mix the gene and vector together in a solution containing the enzyme DNA ligase. This encourages hydrogen bonding between the complementary nucleotides of the gene and plasmid.
-You have now created recombinant DNA which can be inserted in to which ever organism you which to modify.