Related Experiment Video
Updated: Jun 7, 2025

3' End Sequencing Library Preparation with A-seq2
Published on: October 10, 2017
RNA Sequencing Protocols for Short-Read Sequencing
Laura Vasquez-Velez1, Veera D'Mello1, Patricia Soteropoulos2
1Genomics Center, Rutgers New Jersey Medical School, Newark, NJ, USA.
Abstract:
RNA sequencing (RNA-seq) methodologies allow the discovery of novel variants and transcripts. These comprise three general steps: (1) capture of RNA species of interest, (2) conversion of RNA to complementary DNA (cDNA), and (3) modification of cDNA to fit the sequencing platform. Here we describe four different library preparation protocols for short-read sequencing: cDNA synthesis with poly(A) selection, library preparation with ribosomal depletion, and cDNA synthesis with SMART® (Switching Mechanism at 5' end of RNA Template) technology for low and Pico inputs.
Related Concept Videos
RNA-seq
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
Next-generation Sequencing
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
Sanger Sequencing

