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An Efficient Method for the Isolation of Highly Purified RNA from Seeds for Use in Quantitative Transcriptome Analysis
Published on: January 11, 2017
DeLTa-Seq: High-Throughput Targeted RNA-Seq of Rice Leaves Without RNA Purification
Makoto Kashima1, Yasuyuki Nomura2, Atsushi J Nagano3,4
1Faculty of Science, Department of Molecular Biology, Toho University, Chiba, Japan.
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.

