BioTrek Competition
The BioTrek Society was first formed in the 2000-2001 academic year as a pilot project, by Mr. Melegos at GL. Roberts CVI. The BioTrek Society was formed with few student members, and few projects. Currently, membership capacity is limited between 10 to 15 students. During the 2005-2006 academic year The BioTrek Society was relocated at Dunbarton High School.
Its mission is to promote awareness in the subject of applied biotechnology, and to a lesser extent space science. We devote particular emphasis to basic research and open-ended hands-on experiments, through informal seminars and meetings. Numeracy and literacy skills are highly emphasized and applied fully through guided inquiry, and authentic education. The BioTrek Society strives to develop a collegial atmosphere, where high school students are given leadership opportunities to conduct studies (typically performed at the post-secondary level), and disseminate their outcome to peers.
Abstract
DNA methylation is one of several epigenetic mechanisms that can affect an organism’s operation. The purpose of this research was to discover and refine a method to amplify DNA while conserving its methylation pattern. This amplified and methylated DNA could then be transformed into organisms to explore how methylated DNA segments operate and affect gene expression. In order to preserve methylation sites during PCR, we plan to use DNA methyltransferase 1 (DNMT1), the enzyme responsible for conserving methylation in living cells. However, the denaturing temperature for DNMT1 is lower than the temperature reached in standard PCR. For this reason, we have decided to utilize helicase-dependent amplification, an alternative DNA amplification procedure. We are going to combine DNMT1 with Helicase PCR, so that the DNA can be amplified and methylated at the same time. Helicase-dependent amplification operates at lower temperatures than standard PCR, making it suitable for use with DNMT1. Presently, we are working on designing primers suitable for Helicase PCR. This strand will include a promoter/RBS site, red fluorescing protein (RFP) section, and a terminator. We can use this chain to detect whether or not we performed Helicase PCR correctly. Our final DNA strand will also require two restriction cut sites, which will help us to ascertain whether it has been methylated or hemimethylated.
Keywords: DNA Methylation, PCR, DNMT1, Helicase-dependent Amplification, Epigenetics