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复杂植物组织中组蛋白修饰的ChIP-Seq样本与文库制备的高效内部偶联

Efficient In-house Coupling of Sample and Library Preparation for ChIP-Seq of Histone Modifications in Complex Plant Tissues.

作者信息

Long Jiaxin, Johnson Emily T, Ogas Joe

机构信息

Department of Biochemistry, Purdue University, West Lafayette, IN, USA.

出版信息

Methods Mol Biol. 2025;2919:179-198. doi: 10.1007/978-1-0716-4486-7_10.

Abstract

ChIP-seq is a commonly used method to characterize chromatin-associated features on a genome-wide basis. A common set of steps for a ChIP-seq procedure prior to sequencing includes crosslinking, nuclei extraction, chromatin shearing, immunoprecipitation, elution, reversal of crosslinks, and library preparation. Plant material can be challenging to process for ChIP-seq analysis due to the unique attributes of plant cells that impair success. Here, we describe an effective ChIP-seq sample preparation method that is optimized for generating robust libraries from complex plant materials. In particular, we identify time as a critical parameter for effective coupling of ChIP-seq sample preparation with a commercially available kit to generate robust NGS libraries in-house. The resulting protocol is a cost-effective strategy to generate reliable ChIP-seq libraries from complex plant material and thereby acquire representative sequencing data.

摘要

染色质免疫沉淀测序(ChIP-seq)是一种常用于在全基因组范围内表征与染色质相关特征的方法。测序前ChIP-seq程序的一组常见步骤包括交联、细胞核提取、染色质剪切、免疫沉淀、洗脱、交联逆转和文库制备。由于植物细胞的独特属性会影响ChIP-seq分析的成功率,因此处理植物材料进行ChIP-seq分析具有挑战性。在此,我们描述了一种有效的ChIP-seq样本制备方法,该方法经过优化,可从复杂的植物材料中生成可靠的文库。特别是,我们确定时间是将ChIP-seq样本制备与市售试剂盒有效结合以在内部生成可靠的二代测序(NGS)文库的关键参数。所得方案是一种经济高效的策略,可从复杂的植物材料中生成可靠的ChIP-seq文库,从而获得具有代表性的测序数据。

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