Cloning of a PCR Amplified Gene: Lab 331

The Edvotek MegaCycler

Experimental Objective :

This experiment will involve the amplification of a DNA fragment by Polymerase Chain Reactions and to clone the amplified DNA . Bacterial Transformation will provide evidence of success.

Video Presentation on Genetic Engineering : Video

"PRE-LAB ":

Submit this information in hand-written form, scanned for Classroom or on a Google Doc.

Define :

1. Polylinker

2. CRISPR

3. Annealing

4. Eco R1

5. Transformation ( as related to bacteria and genes)

6. Ligation

7. Research background information on E.coli strain JM109.

8. What is the function of T4 DNA Ligase?

LAB REPORT Questions :

Complete these questions for the final lab report. Submit these in hand-written form on paper from each student.

1. Web Research : Describe how human insulin, used to treat diabetic patients is made by the bacterium, E.coli.

2. Lab Procedure Review: Lab 331 PROCEDURE: Document

Explain in your own words, though using some technical terms; How was a gene amplified and cloned in this experiment?

3. Explain what a "GeneChip" microarray is and what it is used to do.

4. Endonucleases are used to "cut" DNA. What enzymes is used for "pasting" the DNA together?

5. Why is a thin-walled 0.2 mL tube used for PCR ?