Anmelden

Babraham institute

2 ARTICLES PUBLISHED IN JoVE

Genetics

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

Laetitia Chauve^*1, Jérémie Le Pen^*2,3,5, Francesca Hodge¹, Pia Todtenhaupt¹, Laura Biggins¹, Eric A. Miska^2,3,4, Simon Andrews¹, Olivia Casanueva¹

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

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.

Biology

Worm-align and Worm_CP, Two Open-Source Pipelines for Straightening and Quantification of Fluorescence Image Data Obtained from Caenorhabditis elegans

Hanneke Okkenhaug¹, Laetitia Chauve¹, Fatemeh Masoudzadeh¹, Lars Okkenhaug¹, Olivia Casanueva¹

¹Babraham Institute

Worm-align/Worm_CP is a simple FIJI/CellProfiler workflow that can be used to straighten and align Caenorhabditis elegans samples and to score whole-worm image-based assays without the need for prior training steps. We have applied Worm-align/Worm_CP to the quantification of heat-shock induced expression in live animals or lipid droplets in fixed samples.