This grant seeks funding to purchase a shaking incubator for classroom use in biology and AP biology courses at NTHS. This instrument allows the growth of live bacterial cultures for classroom experiments. This incubator is the last foundational piece of a modern molecular biology laboratory that the NTHS biology lab does not possess. Functionally, it agitates growing cultures at set temperature for a set period of time. Agitation is required to grow liquid bacterial cultures, and thus the standard incubators we currently have are not sufficient for liquid culture experiments. This instrument is vital to modern DNA based molecular biology as it allows students to grow specific liquid bacterial cultures for use in laboratory procedures like bacterial transformation and plasmid preparation.
The main goal of this grant is to allow students to be able to participate in molecular biology and biotechnology experiments that require bacterial cultures. Specifically NTHS, and NTHS students have been named as collaborators on a recently funded NSF grant, and listed as authors on scientific poster presentations to do just that.
NTHS biology, through Dr. Steakley has already been funded as a broader impacts collaborator on an NSF grant, NSF proposal #2145811, “Cytonuclear stoichiometry of heterotrophic plants”. This grant was prepared by Joel Sharborough (NTHS class of 2007) who is now a professor at New Mexico Technical University and studies genome evolution through the use of DNA sequencing and bioinformatics. Specifically this grant proposes to perform sequencing the genomes of the non-photosynthetic parasitic plants pinedrop and snowflower (collected from the Tahoe basin, live and in person in NTHS AP biology courses). These are plants that no longer perform photosynthesis, and instead are parasites of other plants in their ecosystems to get nutrients. Thus, there are many interesting questions to ask about what changes that has caused in their now unused chloroplast genomes. It is a fascinating example of how evolution might be able to change populations that enter new niches and abandoned old ones.
This project was the first time that snowflower has had its chloroplast genome fully DNA sequenced (2022-2023). In future years we intend to now work on sequencing for pine drops. NTHS students did get to participate in being the first to examine this new sequence data, and could theoretically become authors on the publication describing this data. The NSF grant described above has already allocated $5000 of funding for the sequencing reagents and sequencing machine costs and this grant funding is ongoing in future year. The dramatic decrease in sequencing cost will allow us a mostly complete genome sequence using new sequencing technology called nanopore sequencing (described here: How nanopore sequencing works). This level, and depth of sequencing for this low cost has only become possible in the last few years.
But, in order to answer those questions and get the sequence data we must first extract DNA from these plants and prepare samples for sequencing at NTHS. NTHS already possess the basic lab supplies (pipets, centrifuges (Excellence in Education 2018-2019 grant cycle), gel electrophoresis equipment, PCR machine (Excellence in Education 2017-2018 grant cycle), and DNA extraction reagents to perform much of the required steps in this grant on site. But we specifically lack the ability to grow bacterial cultures used to generate sequencing library reagents. The alternative option has been to pre-make these reagents at NM-Tech university, which is possible, but obviously not ideal and removes the students from a vital hand-on portion of the sample preparation.
A shaking incubator is a standard part of a modern biology laboratory to grow standard microbial cultures and allows scientists to complete a number of modern molecular biology protocols. Having access to this will allow teachers to complete relevant and engaging labs with classes. Examples include: preparing samples for DNA sequencing of bacteria from Lake Tahoe water samples to potentially identify new bacteria present in the lake, creating DNA samples to use in bacterial transformation activities to creating glowing bacteria, modification of DNA via CRISPR gene editing proteins, among others. This tool will allow science students at NTHS to get real world experience using professional level equipment and doing modern, interesting molecular biology experiments. This is a tremendous help because students are able to contextualize and apply the scientific concepts they are studying in their courses with their own hands. With the transition to the Next Generation Science Standards (NGSS) this is now the expectation and the purchase of this tool will allow NTHS students to keep up with the now increasingly rigorous standards. As the district broadly transitions to NGSS science instruction, students will need to engage with tools and labs like these in order to build 21st century skills. These skills are paramount for success in future science careers, and success on upcoming NGSS-aligned science assessments.