Proposal

Narrative

This proposal seeks funding for two Eppendorf thermal cyclers to perform routine Polymerase Chain Reaction (PCR) in my laboratory as a way of discovering novel genes in the nervous system. Since it was invented in 1983, and earned its inventors a Nobel Prize in Chemistry in 1993, PCR has become a widely used technique to generate millions of copies of a specific DNA sample. With this technique, undergraduate and graduate students will be able to test the expression of hundreds of candidate genes in glia cells in the nervous system of zebrafish embryos and discover new glial markers. Zebrafish possess 26,000 genes and about 80% of human genes associated with a human disease have a counterpart in zebrafish. Students will select candidate genes after reviewing the literature and open-source gene expression databases. They will design primers (short nucleotide sequence) to amplify the DNA of the selected genes by using free online tools and will generate millions of copies of the chosen DNA sequences under 2 hours by PCR using the thermal cyclers. After the PCR step, students will synthetize an RNA probe to detect genes in fixed zebrafish embryos through a classic molecular method called in situ hybridization. I expect my lab to have ~5 undergraduate students each year, mentored by graduate students and myself. At first, students will work under the supervision of an experienced lab member, with the goal of eventually conducting independent research in the lab. This project can also be carried out by high school students, graduate students and postdocs.

Facilities

none required

Hardware Requirements

Two Eppendorf Mastercycler X50s - PCR Thermocycler Catalog number. 6311000010 2 x $11,345

Software Requirements

No software required

Personnel Costs

No personnel cost

Other Costs

No other cost

Timeline

Upon approval of the project, undergraduate and graduate students will be able to start doing experiments as soon as the thermo cyclers are received. The samples and reagents will be readily available. The equipment will be used routinely in the lab space in MC-19 on the Jupiter campus in the long term.

Sustainability

Thermo cyclers require very low maintenance and no service contract. They have an average life span of 15 years. In addition to being used for the project proposed in this application, the thermo cyclers will be used on a weekly basis by undergraduate and graduate students in my lab to perform zebrafish genotyping.

Resource Matching

Consumable required for this project (enzymes, buffer, primers, tubes) will be paid with my funds (start-up money and future grants).

Implementing Organization

Department of Biological sciences, College of Science