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BIO May Exam

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Question:

Describe the procedure to make a recombinant plasmid

Author: Rebeca Lorena Arce Fonseca



Answer:

You extract mRNA de un pancreatic exocrine cell. Using reverse transcriptase you make a DNA copy of it, and sticky ends (G nucleotides) are added. Plasmid is cut open with restriction endonuclease and sticky C nucleotide ends are added to it. The DNA of both pancreatic cell and plasmid fuse through complementary base pairing. Ligase then seals the nicks in the recombinant plasmid and the latter is absorbed by the E. coli cell. E. coli is cultured in a fermenter and finally it starts producing human insulin which is then extracted


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