Related Experiment Video
Updated: Apr 17, 2026

Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA
Published on: October 27, 2011
Preparation of cDNA libraries for high-throughput RNA sequencing analysis of RNA 5' ends
Irina O Vvedenskaya1, Seth R Goldman, Bryce E Nickels
1Department of Genetics and Waksman Institute of Microbiology, Rutgers University, Piscataway, NJ, 08854, USA.
Abstract:
We provide a detailed protocol for preparing cDNA libraries suitable for high-throughput sequencing that are derived specifically from the 5' ends of RNA (5' specific RNA-seq). The protocol describes how cDNA libraries for 5' specific RNA-seq can be tailored to analyze specific classes of RNAs based upon the phosphorylation status of the 5' end. Thus, the analysis of cDNA libraries generated by these methods provides information regarding both the sequence and phosphorylation status of the 5' ends of RNAs. 5' specific RNA-seq can be used to analyze transcription initiation and posttranscriptional processing of RNAs with single base pair resolution on a genome-wide level.
Related Concept Videos
RNA-seq
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
RACE - Rapid Amplification of cDNA Ends

