Digital transcriptome profiling using selective hexamer priming for cDNA synthesis
| Autor: | Jyoti Shah, Jason M. Johnson, Christopher K. Raymond, Ronghua Chen, Christopher D. Armour, Patrick M. Loerch, Stuart Jackson, John Dey, Tomas Babak, John C. Castle, Carol A. Rohl |
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| Rok vydání: | 2009 |
| Předmět: |
DNA
Complementary Polyadenylation Oligonucleotide cDNA library Gene Expression Profiling RNA Brain Cell Biology Computational biology Ribosomal RNA Biology Random hexamer Biochemistry Molecular biology Transcriptome Complementary DNA Humans Cloning Molecular Molecular Biology Biotechnology Gene Library |
| Zdroj: | Nature methods. 6(9) |
| ISSN: | 1548-7105 |
| Popis: | To deplete abundant rRNA from total RNA during cDNA library generation, hexamer primers that perfectly match rRNA are removed. These 'not so randomly primed' cDNA libraries can be generated from small amount of total RNA, preserve strand orientation and are equally enriched in polyadenylated and non-polyadenylated transcripts. We developed a procedure for the preparation of whole transcriptome cDNA libraries depleted of ribosomal RNA from only 1 μg of total RNA. The method relies on a collection of short, computationally selected oligonucleotides, called 'not-so-random' (NSR) primers, to obtain full-length, strand-specific representation of nonribosomal RNA transcripts. In this study we validated the technique by profiling human whole brain and universal human reference RNA using ultra-high-throughput sequencing. |
| Databáze: | OpenAIRE |
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