This detailed volume explores newly-developed methods in PIWI-interacting RNAs (piRNAs) research, methods currently applied to other ncRNAs involved in nuclear regulation which can be used to study piRNAs, and piRNA methods applied in non-classical organisms. It also includes several bioinformatic and biophysical methods related to piRNA studies, consistent with the increasing importance of high-throughput sequencing and computational methods. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, piRNA: Methods and Protocols serves as an ideal guide for researchers seeking to elucidate the numerous mysteries of this area of multicellular biology.

Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Inhalt
Functional Analysis of Individual piRNAs in Aedes aegypti Cells and Embryos Using Antisense Oligonucleotides.- CRISPR-Mediated Genome Engineering in Aedes aegypti.- PIWI-Directed DNA Elimination for Tetrahymena Genetics.- Planarian PIWI-piRNA Interaction Analysis Using Immunoprecipitation and piRNA-Sequencing.- Isolation and Processing of Bovine Oocytes for Small RNA Sequencing.- 3D Imaging and In Situ Hybridization for Uncovering the Functions of MicroRNA in Rice Anther.- Cloning, Sequencing, and Linkage Analysis of piRNAs.- Drosophila Genetic Resources for Elucidating piRNA Pathway.- Generation of Stable Drosophila Ovarian Somatic Cell Lines Using the piggyBac System.- Whole Mount RNA FISH Combined with Immunofluorescence for Analysis of the Telomeric Ribonucleoproteins in the Drosophila Germline.- CRISPR-Mediated Activation of Transposable Elements in Embryonic Stem Cells.- Method for Evaluating Effects of Non-Coding RNAs on Nucleosome Stability.- Revisiting the Glass Treatmentfor Single Molecule Analysis of ncRNA Function.- Low Input Genome-Wide DNA Methylation Analysis with Minimal Library Amplification.- Solid-Support Directional (SSD) RNA-Seq as a Companion Method to CLIP-Seq.- UPA-Seq-Based Search Method for Functional lncRNA Candidates.- Large-Scale Analysis of RNA-Protein Interactions for Functional RNA Motif Discovery Using FOREST.- Computational Methods for the Discovery and Annotation of Viral Integrations.- Bioinformatics Approaches for Determining the Functional Impact of Repetitive Elements on Non-Coding RNAs.- Extending and Running the Mosquito Small RNA Genomics Resource Pipeline.- Preparation of Non-Overlapping Transposable Elements (TE) Annotation by Interval Tree.- Statistical Thermodynamics Approach for Intracellular Phase Separation. <p