MAY 10, 2018 10:30 AM PDT

Cost Effective, Robust Digital Gene Expression Profiling of Up to 96 Targets in 96 Samples for Cell Line Screening with nCounter® PlexSet™ Reagents

Sponsored by: NanoString Technologies
Speaker
  • Senior Scientist at NanoString Technologies, Inc.
    Biography
      Dr. Chris Merritt is a molecular biologist with expertise in developmental biology and cancer research. Currently, he is the lead R&D scientist for developing new multiplexing technologies and lyse-n-go optimized protocols for gene expression analysis, along with many other novel technologies at NanoString. He received his Ph.D. in Molecular Biology and Genetics from the Johns Hopkins School of Medicine where he studied gene expression in the C. elegans germline.

    Abstract

    Cell line screening studies require highly efficient protocols for studying many samples in parallel.  We have developed a lyse-and-go protocol for digital gene expression profiling of up to 96-samples by 96-genes in parallel. This protocol is based on nCounter PlexSet Reagents which enable multiplexing of samples and gene targets.  

    The PlexSet “lyse-n-go” protocol is simple and fast and can be divided into 3 steps:  1) lyse cells, 2) set-up hybridization with crude lysate; 3) initiate automated post-hybridization processing and digital counting. It does not require RNA purification or cDNA conversion steps, thus saving time and resources for the researcher. 
    The data presented are crude cells lysates from various primary cell types, two suspension cell lines, and two adherent cell lines using the IO Lymphocyte Activity Panel to measure the gene expression of 5 housekeeper genes and 91 lymphocyte activity genes (96 genes in total). The data demonstrates that up to 96 high-quality, digital data points for gene expression in each of 96 samples can be generated from crude cell lysates in an efficient, high-throughput protocol and that the cell lysate data correlates well with purified total RNA. 


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