APh162 DNA Science Lab
           HOME | WEEK 1 | WEEK 2 | WEEK 3
                                                                                                Eileen Fong, Jin-Hong Kim, Alexander Lin


Just in the past two decades, the technological advances in molecular genetics has been astounding, providing scientists with a wide range of tools to explore fundamental biological sciences.  The use of plasmids, automated PCR, and restriction enzymes allow biologists to create an infinite number of modular vectors that can be designed to explore specific molecular processes.  The genetic switch is one such construct that has been of particular focus for biophysicists giving quantitative results that can be used to study the stochastic nature of gene expression and regulation.  These engineered genes can be combined with fluorescent labelling techniques and advanced microscopy that provide a measure of gene expression down to the single cell level.

The goal of this project is to provide biophysics students with hands-on experience with the  DNA science techniques often described in a majority of biological studies.  Specifically, we will extract the lacZ gene from wild-type E. Coli cells and transform the cells with a pZE21 vector that is kanamycin-resistance and GFP controlled by tetracyclin (ATC) so that we can measure and compare induction.  As a result of this project, students will be familiar with techniques such as:

  • restriction digests

  • polymerase chain reactions

  • gel electrophoresis

  • vector ligation

  • cell transformation

  • electroporation

  • fluorescence microscopy

  • image analysis

The project is split into three weeks:

Week 1: Restriction Digest and Electrophoresis

Week 2: Extraction, Ligation and Transformation

Week 3: Single cell regulation of gene expression

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