WORKSHEET 3 (PCR AND HYBRIDIZATION)

WORKSHEET 3

PCR AND HYBRIDAIZATION

RECOMBINANT DNA TECHNOLOGY (CHAPTER 1)

1. Name the special DNA polymerase used in PCR reactions. Why do we use it? What are the three basic steps of a PCR cycle? Using a single template molecule, how many DNA molecules are generated after 10 cycles of amplification?

2. For a PCR reaction, the following components are added; double starnded DNA (to be amplified), Taq polymerase, NTPs, and double stranded primer in a suitable buffer. However after several cycles in a PCR machine, amplifications was low. Why?

3.With a diagram, depict the Southern Hybridization technique. What is this technique used for?

4. Draw a labelled diagram showing the technique which can be used to identify and locate a specific sequence in a DNA gel using a probe.

5. In Southern blotting technique, why do we bake nitrocellulose membrane after blotting? Why this technique is called blotting technique? Name a technique used to locate RNA in a gel.