Recombinant DNA Technology Problem Set

Problem 2: Recombinant DNA 1

Tutorial to help answer the question

Which of the following tools of recombinant DNA technology is INCORRECTLY paired with its use?

A. restriction endonuclease - production of DNA fragments for gene cloning.
B. DNA ligase - enzyme that cuts DNA, creating sticky ends.
C. DNA polymerase - copies DNA sequences in the polymerase chain reaction.
D. reverse transcriptase - production of cDNA from mRNA.
E. electrophoresis - RLFP analysis.


Creating Recombinant DNA

  • A plasmid vector is digested with EcoRI at a single site to produce two sticky ends.
  • A sample of human DNA is also digested with EcoRI to produce pieces with the same sticky ends.
  • Human DNA- or cDNA copied from mRNA using reverse transcriptase from retroviruses.
  • The two samples are mixed and allowed to hybridize, some molecules will form with pieces of human DNA inserted into the plasmid vector at the EcoRI site.
  • DNA ligase is used to covalently link the fragments.

  • The Biology Project
    University of Arizona
    Updated: July 15, 1999
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