UNIT 4.13 RASL-seq for Massively Parallel and Quantitative Analysis of Gene Expression
Published Online: 1 APR 2012
Copyright © 2012 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Li, H., Qiu, J. and Fu, X.-D. 2012. RASL-seq for Massively Parallel and Quantitative Analysis of Gene Expression. Current Protocols in Molecular Biology. 98:II:4.13:4.13.1–4.13.9.
- Published Online: 1 APR 2012
- Published Print: APR 2012
Large-scale, quantitative analysis of gene expression can be accomplished by microarray or RNA-seq analysis. While these methods are applicable to genome-wide analysis, it is often desirable to quantify expression of a more limited set of genes in hundreds, thousands, or even tens of thousands of biological samples. For example, some studies may require monitoring a sizable panel of key genes under many different experimental conditions, during development, or following treatment with a large library of small molecules, for which current genome-wide methods are either inefficient or cost-prohibitive. This unit presents a method that permits quantitative profiling of several hundred selected genes in a large number of samples by coupling RNA-mediated oligonucleotide Annealing, Selection, and Ligation with Next-Gen sequencing (RASL-seq). The method even allows direct analysis of RNA levels in cell lysates and is also adaptable to full automation, making it ideal for large-scale analysis of multiple biological pathways or regulatory gene networks in the context of systematic genetic or chemical genetic perturbations. Curr. Protoc. Mol. Biol. 98:4.13.1-4.13.9. © 2012 by John Wiley & Sons, Inc.
- gene expression;
- RNA-mediated oligonucleotide;
- bar-coding strategies;
- high-throughput screening