Dilute-’N’-Go sequencing

Prof. Larry Wangh and his lab are interested in detecting changes in mitochondrial genomic sequences that result from aging, disease, or drugs.  To do this, they use LATE-PCR, an advanced form of asymmetric PCR, to detect mutations in the mitochondria by using multiplexes to study many mitochondrial genes at the same time.  LATE-PCR generates single DNA strands that are easily diluted for sequencing.  In the past. they have only been able to sequence one DNA strand from these multiplex reactions.

In a recent publication in Nucleic Acid Research, staff members Yanwei Jia and John Rice, along with Molecular and Cell Biology grad student Adam Osborne, describe the development of a blocking reagent that allows them to sequence both strands of the product DNA, thus allowing for the easy verification of mutations.

The figure at right shows that without a blocker (BLK), one is not able to obtain the excess (XP) strand sequence from a multiplex reaction.  Using a blocker one is able to get not only the limiting (LP) strand, but also the excess strand from the same multiplex

Recent Grant Awards

Neuroscience Ph.D. candidate Melanie Gainey received an NRSA Fellowship from NINDS. Working in the Turrigiano lab, Melanie plans to study the role of the AMPA receptor subunit GluR2 in synaptic scaling in cultural neurons and in vivo using a conditional GluR2 knockout mouse.

Assistant Professor Suzanne Paradis received a Smith Family New Investigator Award from the Richard & Susan Smith Family Foundation. $300,000 in support over three years will support the lab’s efforts to study synapse development and specifically the role of the Sema4B protein in controlling synapse formation.

Professor Leslie Griffith received $1.1 million over 5 years from NIMH to study why sleep is required for effective memory formation. To understand this linkage at a cellular and molecular level, the Griffith lab is defining the circuits that regulate sleep in Drosophila and how these circuits affect memory formation.

Professor Larry Wangh received $1.38 million over the next year from Smiths Detection to continue research and invention of LATE-PCR et al., platform technologies for highly informative detection and diagnosis of nucleic acids in a single tube.  There are ongoing projects looking at applications to cancer, prenatal genetics, and several infectious diseases in people and animals.

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