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SELECTBIO Conferences NGS, SCA, SMA & Mass Spec: Research to Diagnostics 2016

Juan Santiago's Biography

Juan Santiago, Professor, Stanford University

Juan Santiago received his MS and PhD in Mechanical Engineering from the University of Illinois at Urbana-Champaign in 1992 and 1995. His research includes the development of microsystems for on-chip chemical analysis, drug delivery, sample preparation methods, and desalination of water. Applications of this work include molecular medical diagnostics, drug discovery, environmental monitoring, and the production of drinking water. He is a Fellow of the American Physical Society, a Fellow of the American Society of Mechanical Engineering, Fellow of the American Institute for Medical and Biological Engineering, an Associate Editor of the journal Microfluidics and Nanofluidics, co-founder of several companies in the microfluidics area, co-inventor of micron-resolution particle image velocimetry, and director of the Stanford Microfluidics Laboratory. He served as Associate Editor of Lab on a Chip '08-'13. He has given over 30 keynote and named lectures and more than 100 additional invited lectures. As one measure of impact, his work is cited about 1500 times per year. He has graduated 27 PhD students and advised eight postdoctoral researchers. 17 of his former advisees are now professors at major universities. He has authored and co-authored over 150 archival publications and 200 conference papers, and holds 47 patents (24 of which are currently licensed).

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Fractionation and Analysis of Nuclear versus Cytoplasmic Nucleic Acids from Single Cells

Tuesday, 27 September 2016 at 17:35

Add to Calendar ▼2016-09-27 17:35:002016-09-27 18:35:00Europe/LondonFractionation and Analysis of Nuclear versus Cytoplasmic Nucleic Acids from Single CellsNGS, SCA, SMA and Mass Spec: Research to Diagnostics 2016 in San Diego, California, USASan Diego, California,

Single cell analyses (SCA) have become powerful tools in the study heterogeneous cell populations such as tumors and developing embryos.  However, fractionating and analyzing nuclear versus cytoplasmic fractions of nucleic acids remains a challenge as these fractions easily cross-contaminate.  We present a novel microfluidic system that can fractionate and deliver nucleic acid (NA) fractions from the nucleus (nNA) versus the cytoplasm (cNA) from single cells to independent downstream analyses.  Our technique leverages a selective electrical lysis which disrupts the cell’s (outer) cytoplasmic membrane, while leaving the nucleus relatively intact. We selectively extract, purify, and preconcentrate cNA using isotachophoresis (ITP).  The ITP-focused cNA and nNA-containing nucleus are separated by ITP and fractionated at a bifurcation downstream and then extracted for off chip analyses. We will present example applications of this fractionation including qPCR and next generation sequencing (NGS) analyses of cNA vs. nNA.  This will include preliminary NGS analyses of nuclear vs. cytoplasmic RNA fractions to analyze gene expression and splicing.  We hypothesize that the robust and precise nature of our electric field control is amenable to further automation to increase throughput while removing manuals steps.

Add to Calendar ▼2016-09-26 00:00:002016-09-28 00:00:00Europe/LondonNGS, SCA, SMA and Mass Spec: Research to Diagnostics 2016NGS, SCA, SMA and Mass Spec: Research to Diagnostics 2016 in San Diego, California, USASan Diego, California,