Gel-seq: A Method for Simultaneous Sequencing Library Preparation of DNA and RNA Using Hydrogel Matrices.

Autor:	Hoople GD; Department of Engineering, University of San Diego; ghoople@sandiego.edu., Richards A; Department of Bioengineering, University of California, San Diego., Wu Y; Department of Bioengineering, University of California, San Diego., Pisano AP; Department of Mechanical and Aerospace Engineering, University of California, San Diego., Zhang K; Department of Bioengineering, University of California, San Diego.
Jazyk:	angličtina
Zdroj:	Journal of visualized experiments : JoVE [J Vis Exp] 2018 Mar 26 (133). Date of Electronic Publication: 2018 Mar 26.
DOI:	10.3791/57315
Abstrakt:	The ability to amplify and sequence either DNA or RNA from small starting samples has only been achieved in the last five years. Unfortunately, the standard protocols for generating genomic or transcriptomic libraries are incompatible and researchers must choose whether to sequence DNA or RNA for a particular sample. Gel-seq solves this problem by enabling researchers to simultaneously prepare libraries for both DNA and RNA starting with 100 - 1000 cells using a simple hydrogel device. This paper presents a detailed approach for the fabrication of the device as well as the biological protocol to generate paired libraries. We designed Gel-seq so that it could be easily implemented by other researchers; many genetics labs already have the necessary equipment to reproduce the Gel-seq device fabrication. Our protocol employs commonly-used kits for both whole-transcript amplification (WTA) and library preparation, which are also likely to be familiar to researchers already versed in generating genomic and transcriptomic libraries. Our approach allows researchers to bring to bear the power of both DNA and RNA sequencing on a single sample without splitting and with negligible added cost.
Databáze:	MEDLINE
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