DeLTa-Seq：无需 RNA 纯化的水稻叶片高通量靶向 RNA-Seq。

DeLTa-Seq: High-Throughput Targeted RNA-Seq of Rice Leaves Without RNA Purification.

机构信息

Faculty of Science, Department of Molecular Biology, Toho University, Chiba, Japan.

Research Institute for Food and Agriculture, Ryukoku University, Shiga, Japan.

出版信息

Methods Mol Biol. 2025;2869:113-121. doi: 10.1007/978-1-0716-4204-7_12.

DOI:10.1007/978-1-0716-4204-7_12

PMID:39499472

Abstract

DeLTa-seq is a high-throughput RNA-seq library preparation method that enables quantification of the expression of hundreds of arbitrarily selected genes without RNA purification. This method involves direct reverse transcription using rice leaf lysate and targeted RNA-seq library preparation. DeLTa-seq enables the precise quantification of gene expression with a small number of sequencing reads. This chapter provides detailed information on the design of gene-specific primers, sampling of rice leaves, preparation of lysates, direct-lysate reverse transcription, targeted RNA-seq library preparation, and bioinformatic analysis of DeLTa-seq data.

摘要

DeLTa-seq 是一种高通量 RNA-seq 文库制备方法，无需 RNA 纯化即可实现数百个任意选择基因的表达定量。该方法涉及使用水稻叶片裂解物直接进行反转录和靶向 RNA-seq 文库制备。DeLTa-seq 可以用少量测序reads 实现基因表达的精确定量。本章提供了关于基因特异性引物设计、水稻叶片取样、裂解物制备、直接裂解物反转录、靶向 RNA-seq 文库制备以及 DeLTa-seq 数据的生物信息学分析的详细信息。