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imported>Xenbase /* External links to Cold Spring Harbor Xenopus Protocols [http://cshprotocols.cshlp.org/search?fulltext=Xenopus&submit=yes&x=18&y=12&tocsectionid=protocol&tocsectionid=recipe&tocsectionid=topic+introduction&tocsectionid=emerging+model+organisms&tocsec... |
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**Dissection of Xenopus laevis Neural Crest for in vitro Explant Culture or in vivo Transplantation - Millet & Monsoro-Burq [https://www.jove.com/video/51118/dissection-xenopus-laevis-neural-crest-for-vitro-explant-culture-or] | **Dissection of Xenopus laevis Neural Crest for in vitro Explant Culture or in vivo Transplantation - Millet & Monsoro-Burq [https://www.jove.com/video/51118/dissection-xenopus-laevis-neural-crest-for-vitro-explant-culture-or] | ||
=== '''External links to Cold Spring Harbor Xenopus Protocols''' [http://cshprotocols.cshlp.org/search?fulltext=Xenopus&submit=yes&x=18&y=12&tocsectionid=protocol&tocsectionid=recipe&tocsectionid=topic+introduction&tocsectionid=emerging+model+organisms&tocsectionid=product+protocol&tocsectionid=kit&tocsectionid=information+panel] - subscription required, | === '''External links to Cold Spring Harbor Xenopus Protocols''' [http://cshprotocols.cshlp.org/search?fulltext=Xenopus&submit=yes&x=18&y=12&tocsectionid=protocol&tocsectionid=recipe&tocsectionid=topic+introduction&tocsectionid=emerging+model+organisms&tocsectionid=product+protocol&tocsectionid=kit&tocsectionid=information+panel] - subscription required, 2020 articles listed above, earlier protocols below=== | ||
**Raising Antibodies for Use in ''Xenopus''. Maya Z. Piccinni and Matthew J. Guille. [http://cshprotocols.cshlp.org/content/early/2020/01/03/pdb.prot105585.full.pdf+html?sid=972ba2c9-9b73-4306-a49c-bfa681762184] | |||
**Purifying Antibodies Raised against ''Xenopus'' Peptides. Maya Z. Piccinni and Matthew J. Guille. [http://cshprotocols.cshlp.org/content/early/2020/01/03/pdb.prot105619.full.pdf+html?sid=972ba2c9-9b73-4306-a49c-bfa681762184] | |||
**Assessing the Immune Response When Raising Antibodies for Use in ''Xenopus''. Maya Z. Piccinni and Matthew J. Guille. [http://cshprotocols.cshlp.org/content/early/2020/01/03/pdb.prot105593.full.pdf+html?sid=972ba2c9-9b73-4306-a49c-bfa681762184]. | |||
**In Vitro Transcription Systems. Michael R. Green and Joseph Sambrook. [http://cshprotocols.cshlp.org/content/2020/1/pdb.top100750.full.pdf+html?sid=972ba2c9-9b73-4306-a49c-bfa681762184] | |||
**Applying Tensile and Compressive Force to Xenopus Animal Cap Tissue. | |||
**Housing and Feeding of Xenopus laevis - Sive et al. [http://cshprotocols.cshlp.org/content/2007/5/pdb.top8.abstract?sid=009b88bf-7bfb-4279-9528-70643a25bbae] | **Housing and Feeding of Xenopus laevis - Sive et al. [http://cshprotocols.cshlp.org/content/2007/5/pdb.top8.abstract?sid=009b88bf-7bfb-4279-9528-70643a25bbae] | ||
**Inducing Ovulation in Xenopus laevis - Sive et al. [http://cshprotocols.cshlp.org/content/2007/5/pdb.prot4734.abstract?sid=009b88bf-7bfb-4279-9528-70643a25bbae] | **Inducing Ovulation in Xenopus laevis - Sive et al. [http://cshprotocols.cshlp.org/content/2007/5/pdb.prot4734.abstract?sid=009b88bf-7bfb-4279-9528-70643a25bbae] | ||
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**Imaging Axon Pathfinding in Xenopus In Vivo - Leung & Holt [http://cshprotocols.cshlp.org/content/2012/9/pdb.prot070003.abstract?sid=47d75b27-9727-4811-8a78-0330a022b90f] | **Imaging Axon Pathfinding in Xenopus In Vivo - Leung & Holt [http://cshprotocols.cshlp.org/content/2012/9/pdb.prot070003.abstract?sid=47d75b27-9727-4811-8a78-0330a022b90f] | ||
**A Versatile Protocol for mRNA Electroporation of Xenopus laevis Embryos - Chernet & Levin [http://cshprotocols.cshlp.org/content/2012/4/pdb.prot067694.abstract?sid=47d75b27-9727-4811-8a78-0330a022b90f] | **A Versatile Protocol for mRNA Electroporation of Xenopus laevis Embryos - Chernet & Levin [http://cshprotocols.cshlp.org/content/2012/4/pdb.prot067694.abstract?sid=47d75b27-9727-4811-8a78-0330a022b90f] | ||
== '''''General Research Protocols''''' == | == '''''General Research Protocols''''' == |
Revision as of 09:56, 22 January 2020
Books for Xenopus Research and Protocols
- Xenopus Protocols: Post-Genomic Approaches. Hoppler and Vize, 2012 [1]
- expanded second edition with novel approaches inspired by X. tropicalis genome sequencing.
- The Laboratory Xenopus sp. Green, 2010 [2]
- a highly detailed manual containing Xenopus husbandry, management, veterinary care, and frog and equipment vendor information
- Xenopus Protocols: Cell Biology and Signal Transduction Liu (First edition), 2006 [3]
- step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents.
- Color Atlas of Xenopus laevis Histology, Wiechmann and Wirsig-Wiechmann, 2003 [4]
- central source on the microscopic anatomy of cells, tissues, and major organs of Xenopus laevis.
- Early Development of Xenopus laevis: A Laboratory Manual, Sive, Grainger, and Harland, 2000 [5]
- comprehensive collection of protocols for the study of early development in Xenopus embryos
- Transgenic Xenopus: Microinjection Methods and Developmental Neurobiology, Seidman and Soreq, 1997 [6]
- referenced guide to the use of microinjected embryos studying the role and regulation of nervous system proteins during development.
- Atlas of Xenopus Development, Bernardini, Prati, Bonetti, and Scari, 1999 [7]
- scanning, transmission, and light microscopy images of Xenopus embryonic development.
- Normal Table of Xenopus laevis (Daudin), Nieuwkoop and Faber (Third edition), 1994 [8]
- a systematic and chronological description of Xenopus laevis development.
- Xenopus laevis: Practical Uses in Cell and Molecular Biology, Volume 36, Kay and Peng, 1991 [9]
- detailed description of a wide range of uses for Xenopus laevis oocytes and embryos in cell and molecular biology.
- The early development of Xenopus laevis. An atlas of the histology, Hausen and Riebesell, 1991 [10]
- detailed histological sections of Xenopus embryonic development
Online Xenopus Resources
External links to the Journal of Visualized Experiments (JOVE) - Xenopus embryo [11] - excellent video demonstrations
- Functional Evaluation of Olfactory Pathways in Living Xenopus Tadpoles - Terni et al. -[12]
- Obtaining Eggs from Xenopus laevis Females - Cross & Powers [13]
- Microinjection of Xenopus Laevis Oocytes - Cohen et al. - [14]
- Fertilization of Xenopus oocytes, Host Transfer Method - Schneider et al. [15]
- Tissue Determination Using Animal Cap Transplant (ACT) Assay in X. laevis - Viczian & Zuber [16]
- Blastomere Explants to Test for Cell Fate Commitment - Grant et al. [17]
- Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation - Amaya & Kroll [18]
- Organizer and Animal Pole Explants from X. laevis Embryos and Cell Adhesion Assay - Ogata & Cho [19]
- Plastic Embedding and Sectioning of X. laevis Embryos - Ogata1 et al. [20]
- Visualizing RNA Localization in Xenopus Oocytes - Gagnon & Mowry [21]
- Neural Explant Cultures from Xenopus laevis - Lowery et al. [22]
- Translocation of Fluorescent Proteins in Xenopus Ectoderm in Response to Wnt Signaling - Itoh & Sokol [23]
- Dissection, Culture, and Analysis of X. laevis Embryonic Retinal Tissue - McDonough et al. [24]
- X. tropicalis Egg Extracts to Identify Microtubule-associated RNAs - Sharp & Blower [25]
- Electroporation of Craniofacial Mesenchyme - Tabler & Liu [26]
- Electrophysiological Recording from Xenopus Nerve-muscle Co-cultures - Yazejian et al. [27]
- Single Cell Electroporation in vivo within the Intact Developing Brain - Hewapathirane & Haas - [28]
- Live-cell Imaging and Quantitative Analysis of Embryonic Epithelial Cells - Joshi & Davidson [29]
- Preparation and Fractionation of Xenopus laevis Egg Extracts - Cross & Powers [30]
- In Vitro Nuclear Assembly Using Fractionated Xenopus Egg Extracts - Cross & Powers [31]
- Study of the DNA Damage Checkpoint using Xenopus Egg Extracts - Willis et al. [32]
- Two Types of Assays for Detecting Frog Sperm Chemoattraction - Burnett et al. [33]
- Comparative in vivo Study of gp96 Adjuvanticity in X. laevis - Nedelkovska et al. [34]
- Patch Clamp and Perfusion Techniques - Yang et al. [35]
- Patch Clamp Recording of Ion Channels - Brown et al. [36]
- Cation Transport in Xenopus Oocytes - Dürr et al.[37]
- Stem cell-like Xenopus Embryonic Explants to Study Early Neural Developmental Features In Vitro and In Vivo - BC.Durand [38]
- Dissection of Xenopus laevis Neural Crest for in vitro Explant Culture or in vivo Transplantation - Millet & Monsoro-Burq [39]
External links to Cold Spring Harbor Xenopus Protocols [40] - subscription required, 2020 articles listed above, earlier protocols below
- Raising Antibodies for Use in Xenopus. Maya Z. Piccinni and Matthew J. Guille. [41]
- Purifying Antibodies Raised against Xenopus Peptides. Maya Z. Piccinni and Matthew J. Guille. [42]
- Assessing the Immune Response When Raising Antibodies for Use in Xenopus. Maya Z. Piccinni and Matthew J. Guille. [43].
- In Vitro Transcription Systems. Michael R. Green and Joseph Sambrook. [44]
- Applying Tensile and Compressive Force to Xenopus Animal Cap Tissue.
- Housing and Feeding of Xenopus laevis - Sive et al. [45]
- Inducing Ovulation in Xenopus laevis - Sive et al. [46]
- Xenopus laevis In Vitro Fertilization and Natural Mating Methods - Sive et al. [47]
- Egg Collection and In Vitro Fertilization of the Western Clawed Frog Xenopus tropicalis - Showell & Conlon [48]
- Isolation of Xenopus Oocytes - Sive et al. [49]
- Isolating Xenopus laevis Testes - Sive et al. [50]
- Dejellying Xenopus laevis Embryos - Sive et al. [51]
- Removing the Vitelline Membrane from Xenopus laevis Embryos - Sive et al. [52]
- Microinjection of Xenopus Embryos - Sive et al. [53]
- Defolliculation of Xenopus Oocytes - Sive et al. [54]
- Microinjection of Xenopus Oocytes - Sive et al. [55]
- Animal Cap Isolation from Xenopus laevis - Sive et al. [56]
- Xenopus laevis Keller Explants - Sive et al. [57]
- Microinjection of RNA and Preparation of Secreted Proteins from Xenopus Oocytes - Sive et al. [58]
- Calibration of the Injection Volume for Microinjection of Xenopus Oocytes and Embryos - Sive et al. [59]
- Isolation of DNA from Red Blood Cells in Xenopus - Sive et al. [60]
- Investigating Morphogenesis in Xenopus Embryos: Imaging Strategies, Processing, and Analysis - Kim & Davidson [61]
- Low-Magnification Live Imaging of Xenopus Embryos for Cell and Developmental Biology - Wallingford [62]
- High-Magnification In Vivo Imaging of Xenopus Embryos for Cell and Developmental Biology - Keiserman et al. [63]
- Preparation of Fixed Xenopus Embryos for Confocal Imaging - Wallingford [64]
- Whole-Mount Fluorescence Immunocytochemistry on Xenopus Embryos - Lee et al. [65]
- Generation of Transgenic Xenopus laevis - Kroll & Amaya - [66] [67] [68]
- In Vivo Time-Lapse Imaging of Neuronal Development in Xenopus - Ruthazer et al. [69]
- Photoconversion for Tracking the Dynamics of Cell Movement in Xenopus laevis Embryos - Chernet et al. [70]
- Single-Cell Electroporation in Xenopus - Liu & Haas [71]
- Imaging Axon Pathfinding in Xenopus In Vivo - Leung & Holt [72]
- A Versatile Protocol for mRNA Electroporation of Xenopus laevis Embryos - Chernet & Levin [73]
General Research Protocols
Animal Husbandry
- obtain frogs and frog housing equipment [74]
- Xenopus Express links [75] - Xenopus Express husbandry [76]
- Xenopus tropicalis and Xenopus laevis environmental parameter standard operating procedure (Kroll lab)
Lab Solutions and Reagents (click each to view expanded content)
- culture media
- immunohistochemistry
- in situ hybridization
- fixatives
- DNA/RNA preparation
- embryo manipulation/dissection
- microinjection
- general buffers and salt solutions
Generating Embryos
- inducing egg laying via hCG injection (Vize lab)
- anesthesia, in vitro fertilization, and natural mating of Xenopus tropicalis (Conlon lab)
- benzocaine stock, 100x (Khokha lab)
- in vitro fertilization of Xenopus laevis (Conlon lab)
- high-salt solution for egg laying (Zorn lab/Ron Laskey)
- dejellying embryos (Sive et al. 2000)
- dejellying embryos (Zorn lab)
Transgenesis
- transgenesis in Xenopus laevis - adapted from Amaya et al 2001 (Conlon lab)
- tadpole tail sampling for genomic DNA (Conlon lab)
- Production of transgenic Xenopus laevis by restriction enzyme mediated integration and nuclear transplantation. Amaya E, Kroll K. J Vis Exp. 2010 Aug 21;(42). [77]
- A method for generating transgenic frog embryos. Ishibashi S, Kroll KL, Amaya E. Methods Mol Biol. 2008;461:447-66. [78]
- A method for generating transgenic frog embryos. Amaya E, Kroll KL. Methods Mol Biol. 1999;97:393-414. [79]
in situ Hybridization
- in situ hybridization, brief version (Harland lab)
- whole-mount in situ hybridization (Conlon lab)
- whole-mount in situ hybridization - LNA probes (Wheeler lab)
- fluorescent in-situs and FCIS (Vize lab)
- RNA Whole-Mount In situ Hybridisation Proximity Ligation Assay (rISH-PLA) (Myers and Scarlett labs)
Immunohistochemistry
- wholemount antibody staining (CSH Protocols) (Wallingford lab)
- wholemount antibody staining - alkaline phosphatase detection (Conlon lab)
- wholemount antibody staining - adapted from Dan Weeks' protocol (Conlon lab)
- immunostaining sectioned embryos (Conlon lab)
- hydrochloric acid antigen retrieval (Zorn lab)
- RNA Whole-Mount In situ Hybridisation Proximity Ligation Assay (rISH-PLA) (Myers and Scarlett labs)
ChIP protocols
- Chromatin immunoprecipitation analysis of Xenopus embryos., Methods Mol Biol. 2012;917:279-92. [80] [81]
- Akkers RC, Jacobi UG, Veenstra GJ.
- Chromatin immunoprecipitation in early Xenopus laevis embryos., Dev Dyn. 2009 Jun;238(6):1422-32. [82] [83]
- Blythe SA, Reid CD, Kessler DS, Klein PS.
Histology
Embryo Staining Protocols (non in situ)
- benzidine-peroxidase staining for blood (Conlon lab)
- benzidine-peroxidase staining for blood (Thomsen lab)
- apoptosis via TUNEL (Veenstra lab)
- TUNEL staining of whole Xenopus embryos - Harland protocol (Conlon lab)
Immuno and Protein Protocols
- immunoprecipitation (Stukenberg lab)
- immunofluorescence with biotinylated antibodies (Stukenberg lab)
- Purifying Polyclonal Antibodies using Affinity Columns/Chromatography
- Making an Affinity Column
- Running an Affinity Column
- whole embryo protein extraction with freon (Conlon lab)
- SDS-PAGE gels (Conlon lab)
- western blot protocol (Conlon lab)
- silver staining protein gels (Stukenberg lab)
Nucleic Acid Protocols
- RNA Isolation
- DNA Isolation
Oocyte Transfer Technique (Heasman/Wylie labs)
- Part 1: Choosing Oligodeoxyribonucleotides (ODNs)
- Part 2: ODN Modification
- Part 3: Host Transfer Technique
- Part 4: Fertilization and Development
Xenopus Oocyte and Egg Extracts
- crude interphase egg extract (Shechter lab)
- isolation of chromatin from egg extract and histone recovery (Shechter lab)
- preparation of cycling egg extract (Shechter lab)
- cycling extracts, mike's protocol (Stukenberg lab)
- cycling extracts, sara's protocol (Stukenberg lab)
- CSF extract, mike's protocol (Stukenberg lab)
- CSF extract, sara's protocol (Stukenberg lab)
- assaying H1 kinase in extracts (Stukenberg lab)
- frozen egg extracts that retain spindle assembly activity (Takagi and Shimamoto, 2017)
Morpholino Studies in Xenopus Brain Development
Bestman and Cline 2019, Brain Development pp 377-395, Part of the Methods in Molecular Biology book series (MIMB, volume 2047) [[84]]