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Preparation of Drosophila Polytene Chromosomes, Followed by Immunofluorescence Analysis of Chromatin Structure by Multi-fluorescence Correlations

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Low-cost and Multiplexable Whole mRNA-Seq Library Preparation Method with Oligo-dT Magnetic Beads for Illumina Sequencing Platforms

Authors: Makoto Kashima, Ayumi Deguchi, Ayumi Tezuka and Atsushi J. Nagano, date: 06/20/2020, view: 900, Q&A: 0

[Abstract] RNA-Seq is a powerful method for transcriptome analysis used in varied field of biology. Although several commercial products and hand-made protocols enable us to prepare RNA-Seq library from total RNA, their cost are still expensive. Here, we established a low-cost and multiplexable whole mRNA-Seq library preparation method for illumine ...

Quantitative Determination of Ca²⁺-binding to Ca²⁺-sensor Proteins by Isothermal Titration Calorimetry

Authors: Seher Abbas and Karl-Wilhelm Koch, date: 04/05/2020, view: 623, Q&A: 0

[Abstract] Diverse and complex molecular recognitions are central elements of signal transduction cascades. The strength and nature of these interaction modes can be determined by different experimental approaches. Among those, Isothermal titration calorimetry (ITC) offers certain advantages by providing binding constants and thermodynamic parameters from ...

mRNA Extraction from Gill Tissue for RNA-sequencing

Authors: Jukka-Pekka Verta and Felicity Jones, date: 03/05/2020, view: 733, Q&A: 0

[Abstract] Adaptation is thought to proceed in part through spatial and temporal changes in gene expression. Fish species such as the threespine stickleback are powerful vertebrate models to study the genetic architecture of adaptive changes in gene expression since divergent adaptation to different environments is common, they are abundant and easy to study ...

Zebrafish Bacterial Infection Assay to Study Host-Pathogen Interactions

Authors: Faiza Basheer, Clifford Liongue and Alister C Ward, date: 03/05/2020, view: 745, Q&A: 0

[Abstract] The study of host-pathogen interactions has improved our understanding of both pathogenesis and the response of the host to infection, including both innate and adaptive responses. Neutrophils and macrophages represent the first line of innate host defense against any infection. The zebrafish is an ideal model to study the response of these cells ...

Retina Injury and Retina Tissue Preparation to Study Regeneration in Zebrafish

Authors: Poonam Sharma and Rajesh Ramachandran, date: 12/20/2019, view: 1237, Q&A: 0

[Abstract] Unlike mammals, primitive vertebrates have immense capability to regenerate almost all of their organs including the central nervous system. Among primitive organisms, zebrafish have been extensively used as a model system for regeneration studies. The retina is a part of the central nervous system and mammals lack the potential to repair any ...

Looking through Brains with Fast Passive CLARITY: Zebrafish, Rodents, Non-human Primates and Humans

Authors: Farzad Mortazavi, Alexander J. Stankiewicz and Irina V. Zhdanova, date: 08/05/2019, view: 2832, Q&A: 0

[Abstract] Recently developed CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging/Immunostaining/In situ-hybridization-compatible Tis-sue-hYdrogel) technique renders the tissue transparent by removing lipids in the tissue, while preserving and stabilizing the cellular and subcellular structures. This provides effective penetration of diverse ...

A Standardized Tank Design for the Light Dark Task in Zebrafish

Authors: Amanda Facciol, Steven Tran and Robert Gerlai, date: 07/20/2019, view: 1769, Q&A: 0

[Abstract] The light dark paradigm is a common behavioral test used to screen a variety of pharmacological agents, including anxiogenics and anxiolytics. Although most often used in rodents, the light dark task has recently been adapted for use in zebrafish. However, a number of inconsistent findings have been reported for this species. Some have found ...

Quantification of Cutaneous Ionocytes in Small Aquatic Organisms

Authors: Garfield T. Kwan, Shane H. Finnerty, Nicholas C. Wegner and Martin Tresguerres, date: 05/05/2019, view: 1690, Q&A: 0

[Abstract] Aquatic organisms have specialized cells called ionocytes that regulate the ionic composition, osmolarity, and acid/base status of internal fluids. In small aquatic organisms such as fishes in their early life stages, ionocytes are typically found on the cutaneous surface and their abundance can change to help cope with various metabolic and ...

Detection of mRNA by Whole Mount in situ Hybridization and DNA Extraction for Genotyping of Zebrafish Embryos

Authors: Rachna Narayanan and Andrew C. Oates, date: 03/20/2019, view: 5996, Q&A: 0

[Abstract] In situ hybridization is used to visualize the spatial distribution of gene transcripts in tissues and in embryos, providing important information about disease and development. Current methods involve the use of complementary riboprobes incorporating non-radioactive labels that can be detected by immunohistochemistry and coupled to ...

Notochord Injury Assays that Stimulate Transcriptional Responses in Zebrafish Larvae

Authors: Zhiqiang Zeng, Juan C Lopez-Baez, Laura Lleras-Forero, Hannah Brunsdon, Cameron Wyatt, Witold Rybski, Nicholas D Hastie, Stefan Schulte-Merker and E Elizabeth Patton, date: 12/05/2018, view: 2160, Q&A: 0

[Abstract] Zebrafish have become an increasingly important model organism in the field of wound healing and regenerative medicine, due to their high regenerative capacity coupled with high-resolution imaging in living animals. In a recent study, we described multiple physical and chemical methods to induce notochord injury that led to highly specific ...

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