0534492967, Barnum, Biotechnology 2e
Chapter 3
objectives
3
1
Basic Principles of Recombinant DNA Technology
Learning Objectives
After studying this chapter in the text, the student should be able to:
1.
Know the function of restriction endonucleases, how they work to cut DNA, and
why they are important in biotechnology. Compare blunt ends with sticky ends.
2.
Know the mechanism by which electrophoresis separates pieces of DNA.
3.
List and know the steps of DNA cloning.
4.
Know how vectors are used to transform bacteria, and know the methods of selecting
for successfully transformed bacteria. Compare the types of vectors in terms of the
sizes of DNA that can be carried by the vector into the bacteria.
5.
List the types of vectors that can be used to transform yeast mammalian cells and
plants, and why they are effective in those organisms.
6.
List the methods of transformation of cells.
7.
Compare genomic libraries, cDNA libraries, and expression libraries in terms of how
they are constructed, what the libraries are looking for, and how they are screened.
8.
List the various types of reporter genes used in research.
9.
Compare Northern and Southern blot hybridization in terms of how they are
constructed and what each type of hybridization is looking for.
10. Know the function of PCR, the steps of PCR, and what PCR allows researchers to
accomplish.
11. Compare the two methods of DNA sequencing: the chemical method and the Sanger
method, and know which method is more widely used. How does automation
impact DNA sequencing?
12. List and define the various methods of analyzing proteins. Are any of these methods
similar to DNA methods?
13. Know the types of microarrays, and how DNA and protein microarrays work.
14. List the applications of recombinant DNA technology.