BrAD-seq: Breath Adapter Directional sequencing: a streamlined, ultra-simple and fast library preparation protocol for strand specific mRNA library construction.
BrAD-seq: Breath Adapter Directional sequencing: a streamlined, ultra-simple and fast library preparation protocol for strand specific mRNA library construction.
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DOI：
10.3389/fpls.2015.00366
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发表时间：
2015
影响因子：
5.6
通讯作者：
Sinha NR
中科院分区：
文献类型：
--
作者：
Townsley BT;Covington MF;Ichihashi Y;Zumstein K;Sinha NR
下一代测序（NGS）推动着生物学认知的快速进步，RNA测序（RNA - seq）已成为生物学和医学不可或缺的工具。尽管NGS文库的制备仍然是其更广泛应用的瓶颈，但对获取这些技术的需求日益增长。在此我们报道一种制备链特异性RNA - seq文库的新方法，该方法利用双链cDNA的末端呼吸作用来捕获并整合测序接头。呼吸接头定向测序（BrAD - seq）减少了样本操作，并且相较于大多数现有方法，其所需的酶促步骤要少得多，却能产生高质量的链特异性RNA - seq文库。我们所提出的方法针对3'端数字基因表达（DGE）文库进行了优化，并且能够轻松扩展到全转录组覆盖的鸟枪法（SHO）类型的链特异性文库，而且该方法是模块化的，可适应多种RNA和DNA输入材料。BrAD - seq为RNA - seq文库提供了一种高度简化且成本低廉的选择。
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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影响因子：
--
作者：
Meyer, Matthias;Kircher, Martin
通讯作者：
Kircher, Martin
影响因子：
5.6
作者：
Goff SA;Vaughn M;McKay S;Lyons E;Stapleton AE;Gessler D;Matasci N;Wang L;Hanlon M;Lenards A;Muir A;Merchant N;Lowry S;Mock S;Helmke M;Kubach A;Narro M;Hopkins N;Micklos D;Hilgert U;Gonzales M;Jordan C;Skidmore E;Dooley R;Cazes J;McLay R;Lu Z;Pasternak S;Koesterke L;Piel WH;Grene R;Noutsos C;Gendler K;Feng X;Tang C;Lent M;Kim SJ;Kvilekval K;Manjunath BS;Tannen V;Stamatakis A;Sanderson M;Welch SM;Cranston KA;Soltis P;Soltis D;O'Meara B;Ane C;Brutnell T;Kleibenstein DJ;White JW;Leebens-Mack J;Donoghue MJ;Spalding EP;Vision TJ;Myers CR;Lowenthal D;Enquist BJ;Boyle B;Akoglu A;Andrews G;Ram S;Ware D;Stein L;Stanzione D
通讯作者：
Stanzione D
影响因子：
2.9
作者：
von Hippel, Peter H.;Johnson, Neil P.;Marcus, Andrew H.
通讯作者：
Marcus, Andrew H.
影响因子：
14.8
作者：
通讯作者：
--
影响因子：
12.3
作者：
Mignone F;Gissi C;Liuni S;Pesole G
通讯作者：
Pesole G