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Genetics

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High-Throughput Quantitative RT-PCR in Single and Bulk C. elegans Samples Using Nanofluidic Technology

Article DOI: 10.3791/61132-v • 08:19 min • May 28th, 2020
May 28th, 2020 •
*1, *2,3,5, 1, 1, 1, 2,3,4, 1, 1
1Babraham Institute, 2Gurdon Institute, University of Cambridge, 3Department of Genetics, University of Cambridge, 4Wellcome Trust Genome Campus, Wellcome Trust Sanger Institute, 5Laboratory of Virology and Infectious Disease, The Rockefeller University
* These authors contributed equally

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Summary
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In this article a high-throughput protocol for fast and reliable determination of gene expression levels in single or bulk C. elegans samples is described. This protocol does not require RNA isolation and produces cDNA directly from samples. It can be used together with high-throughput multiplexed nanofluidic real-time qPCR platforms.

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High-Throughput Quantitative RT-PCR Single Samples Bulk Samples Nanofluidic Technology Throughput Time Efficiency RT-qPCR Data RNA Isolation Bench Work 96-well QPCR Faster Assay Robust Assay Highly Sensitive Assay Gene Expression Variability Isogenic Worms Picking Worms Acclimating Worms Lysis Buffer Dnase I PCR Strip Domed PCR Tube Caps Transferring Worms Liquid Nitrogen Freeze/thaw Procedure
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