(SW4) Bridging the Gap: Isolation to Translation (Single Cell RNA-seq)

Workshop Sponsors:

 

Sunday, April 22

8:00 am – 4:00 pm

Presenters and
Organizers
Mehrnoosh Abshari, National Institutes of Health
Kathleen Brundage, University of West Virginia
Laura Blunk, Stowers Institute for Medical Research
Michael Meyer, University of Pittsburgh Cancer Institute
 
Workshop Overview

RNA-seq has revolutionized transcriptomics and rapidly become the method of choice to address both quantitative and qualitative aspects of gene expression. Single-cell transcriptomics using next-generation transcript sequencing (RNA-seq) is now emerging as a powerful tool to profile cell-to-cell variability on a genomic scale. Its application has already greatly impacted our conceptual understanding of diverse biological processes with broad implications for both basic and clinical research.

The pipeline from original sample to analyzed results involves a variety of cores that are not always up-to-date on each other’s part of the process. Understanding key issues encountered by other cores, as well as how each core’s work affects the others is critical as we seek to better collaborate and troubleshoot experiments. In this workshop, we will review each step of the pipeline (single cell isolation methods, single-cell and bulk RNA-seq protocols and techniques, gene expression data analysis, and applications of RNA-seq) and discuss solutions to facilitate the intersection of multiple cores.

This workshop will be of great value to researchers who are planning experiments that involve the isolation of single cells as well as those planning to perform molecular analysis of single cells purified by flow cytometric cell sorting and other techniques. Core managers, particularly in the fields of Cytometry and Genomics, would specifically benefit, as would post docs and investigators who are experts in one area but would like to have a better understanding of the other pieces of the puzzle for genomic profiling. This workshop will also provide a great networking opportunity for researchers from different disciplines who need to connect and collaborate.

 
Agenda
Introduction
 
Single cell isolation by different techniques
     1. Sample Prep and Flow Sorting
     2. AutoMACs and Magnetic Bead Isolation
Coffee break
     3. Microfluidics-based isolation
     4. Laser capture microdissection
 
Problems, Solutions, and Pitfalls on Single Cell Isolation
(Panel and open discussion)
 
Lunch
 
RNAseq technology
      1. RNAseq
     2. Dropseq
     3. 10X Genomics
 
Coffee Break
 
Bioinformatics 101
 
Applications and Biological significance Potential topics
     1. Stem cell differentiation
     2. Whole tissue analysis
     3. Tumor heterogeneity
 
Conclusion/remarks/Q&A (panel)

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