Rice Methods and Protocols

This volume aims to be a comprehensive guide to the field of rice molecular biology, encompassing both time-honored techniques and state-of-the-art methodologies. Chapters detail sample preparation, temporal and spatial analysis of RNA, protein expression, transformation and precise genome editing, scalable transcriptome analysis, epigenome analysis, and gene regulatory network analysis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Rice: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Hydroponic culture of rice seedlings for stress response assay.- Total RNA extraction from rice vegetative tissues using magnetic beads.- Total RNA extraction from starchy rice endosperm.- RNA extraction from rice immature embryo using laser capture microdissection.- Using immunoblotting for detecting of a circadian clock component, Os-GI, in rice leaves.- Preparation and Sectioning of Paraffin-Embedded Tissue for Histology and Histochemistry.- Spatial Analysis of Gene Expression by In Situ Hybridization.- Purification of Anti-OSH1 IgG and Immunohistochemical Staining of Rice Shoot Apical Meristem.- Agrobacterium-mediated transformation and targeted mutagenesis using SpCas12f in rice.- Gene targeting via CRISPR/Cas9-mediated DNA double-strand break induction in rice.- Precise base substitution using CRISPR/Cas-mediated base editor in rice.- DeLTa-seq: High-throughput targeted RNA-seq of rice leaves without RNA purification.- Lasy-seq: a high-throughput 3' RNA-seq method for large-scale transcriptome profiling in rice.- Profiling of 24nt phasiRNAs in rice anther by massively parallel sequencing and stem-loop reverse transcription PCR.- Expression and purification of pAG-MNase for CUT&RUN.- Chromatin immunoprecipitation from shoot apices and young panicles of rice.- In vitro Genome-wide identification of transcription factor binding sites.- High-efficient transient gene transfection for transcriptional activity assay using rice protoplasts on large-scale platform.