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Next-generation sequencing (NGS) has increased our understanding of biology by enabling highly sensitive RNA expression analysis across a wide dynamic range. As NGS applications continue to grow, so does the need for more powerful tools to work with less-than-ideal samples. As the core of the SMARTer kits for transcriptome analysis, Clontech’s patented SMART® technology utilizes the template switching activity of reverse transcriptase to enable researchers to analyze their most challenging samples, such as single cells, low-input RNA, noncoding RNA, and RNA from degraded samples. In particular, single-cell RNA-seq is one of the more difficult, and fastest growing, applications of NGS. The high sensitivity and dT-primed protocol of the SMARTer Ultra Low family has made these kits the industry standard for single-cell analysis. Applications of SMART technology are constantly expanding and now include a novel, ligation-free method for generating ChIP (chromatin immunoprecipitation) sequencing libraries. The purpose of this seminar is to take a deeper look into the new technologies being developed for single-cell RNA-seq and other sensitive NGS applications.

I will discuss techniques that allow us to perform cellular and subcellular resolution imaging of neuronal activity in mice navigating in virtual reality environments and recent results from imaging place cells and grid cells. I will describe activity patterns that we have observed in hippocampal place cell dendrites and the implications for how associative Hebbian learning may take place during behavior. I will also describe the functional micro-organization of grid cells in the medial entorhinal cortex and what the organization might tell us about the circuits that generate grid cell firing patterns.