Barcoded cDNA library preparation for small RNA profiling by next-generation sequencing

The characterization of post-transcriptional gene regulation by small regulatory (20–30 nt) RNAs, particularly miRNAs and piRNAs, has become a major focus of research in recent years. A prerequisite for characterizing small RNAs is their identification and quantification across different development...

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Published inMethods (San Diego, Calif.) Vol. 58; no. 2; pp. 164 - 170
Main Authors Hafner, Markus, Renwick, Neil, Farazi, Thalia A., Mihailović, Aleksandra, Pena, John T.G., Tuschl, Thomas
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 01.10.2012
Subjects
Base Sequence
Deep sequencing
Digital gene expression
Gene Expression Profiling - methods
Gene Library
High-Throughput Nucleotide Sequencing - methods
Humans
MicroRNAs - chemistry
MicroRNAs - genetics
miRNA
piRNA
RNA, Small Interfering - chemistry
RNA, Small Interfering - genetics
Sequence Analysis, RNA - methods
Specimen Handling
Deep sequencing
miRNA
Digital gene expression
piRNA
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Summary:The characterization of post-transcriptional gene regulation by small regulatory (20–30 nt) RNAs, particularly miRNAs and piRNAs, has become a major focus of research in recent years. A prerequisite for characterizing small RNAs is their identification and quantification across different developmental stages, and in normal and disease tissues, as well as model cell lines. Here we present a step-by-step protocol for generating barcoded small RNA cDNA libraries compatible with Illumina HiSeq sequencing, thereby facilitating miRNA and other small RNA profiling of large sample collections.
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ISSN:1046-2023
1095-9130
DOI:10.1016/j.ymeth.2012.07.030