Journal
FRONTIERS IN PLANT SCIENCE
Volume 6, Issue -, Pages -Publisher
FRONTIERS MEDIA SA
DOI: 10.3389/fpls.2015.00366
Keywords
strand-specific sequencing; NGS; Illumina; RNA-seq libraries; Bioinformatics; BrAD-seq
Categories
Funding
- NSF [IOS-1238243]
- RIKEN Special Postdoctoral Researchers Program
- NIH [S10RR029668, S10RR027303]
- National Science Foundation [DBI-0735191]
- Grants-in-Aid for Scientific Research [15K18589] Funding Source: KAKEN
- Division Of Integrative Organismal Systems
- Direct For Biological Sciences [1238243] Funding Source: National Science Foundation
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Next Generation Sequencing (NGS) is driving rapid advancement in biological understanding and RNA-sequencing (RNA-seq) has become an indispensable tool for biology and medicine. There is a growing need for access to these technologies although preparation of NGS libraries remains a bottleneck to wider adoption. Here we report a novel method for the production of strand specific RNA-seq libraries utilizing the terminal breathing of double-stranded cDNA to capture and incorporate a sequencing adapter. Breath Adapter Directional sequencing (BrAD-seq) reduces sample handling and requires far fewer enzymatic steps than most available methods to produce high quality strand-specific RNA-seq libraries. The method we present is optimized for 3-prime Digital Gene Expression (DGE) libraries and can easily extend to full transcript coverage shotgun (SHO) type strand-specific libraries and is modularized to accommodate a diversity of RNA and DNA input materials. BrAD-seq offers a highly streamlined and inexpensive option for RNA-seq libraries.
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