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Lowering the Limits for Epigenetic Methylation Analysis
This webinar discusses a new library preparation method that enables complete methylome characterization from only about 20 starting cells. A key limitation to epigenetic studies has been the amount of starting material that is required to perform DNA sequencing of bisulfite-converted samples. Swift Biosciences has addressed this with a new method that dramatically reduces the amount of required starting material. The Accel-NGS® Methyl-Seq DNA Library Kit is based on Swift’s Adaptase™ technology, a molecular biology method compatible with single-stranded DNA. Researchers can use Adaptase technology to create next-generation sequencing libraries after bisulfite treatment, thereby generating high-quality DNA sequencing data from five hundredfold less input DNA than other commercially available kits. In this online seminar, Chongyuan Luo of Joseph Ecker’s lab at the Salk Institute details a comparison of the Swift Biosciences method with traditional methylC-seq and also discusses several projects that have pushed the limits of the technology. He shows comparable coverage and strand evenness of a complex methylome library prepared from 1 ng of Arabidopsis DNA as compared to methylC-seq results. Additionally, the lab has successfully prepared “direct” methylome libraries by fragmenting DNA solely with bisulfite conversion and thus removing the need of physical DNA fragmentation. Finally, methylome libraries were prepared from 10 mammalian neuron nuclei by combining bisulfite conversion and DNA purification into one step. This novel approach allows for efficient methylome library preparation from tissues and preserves precious DNA from minute amounts of starting cells.
Chongyuan Luo, Research Associate, Salk Institute of Biological Studies
Chongyuan Luo received his PhD at Rutgers University under the supervision of Eric Lam. During his graduate study, he investigated the functions of chromatin modifications using epigenomic profiling and global correlative analyses. Chongyuan joined Joseph Ecker’s lab at the Salk Institute in May of 2012. He is interested in studying the spatial and temporal dynamics of the epigenome in plants and animals. Currently he is developing methods to enable the profiling of DNA base modifications and other epigenetic marks with minute amounts of starting cells or tissues.
Cassie Schumacher, Research Scientist, Swift Biosciences
Cassie Schumacher holds a BS in biology and biotechnology from DePaul University and an MS in microbiology and immunology from the University of Michigan. In her role as a research scientist at Swift Biosciences, she focuses on development efforts and application support for methylation sequencing as well as low input and difficult to process samples.