Protocols
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]
Protocols published in Journals
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) [[40]]
Cold Spring Harbor Xenopus Protocols Edited by Hazel Sive. published online 2018, 2019 and 2020. [41]
Note: External links are provided but a subscription fee is required for access most CSHL protocols. All CSHL protocol articles are Copyright 2018, 2019, or 2020 Cold Spring Harbor Laboratory Press- Request permission to re-use images/figures accordingly.
General Research Protocols
- Whole-Mount In Situ Hybridization of Xenopus Embryos. Jean-Pierre Saint-Jeannet. 2017 [42]
- Whole-Mount In Situ Hybridization of Xenopus Oocytes. Diana Bauermeister and Tomas Pieler. 2018. [43]
- Fluorescence In Situ Hybridization of Cryosectioned Xenopus Oocytes. Christopher R. Neil and Kimberly Mowry. 2018. [44]
- Whole-Mount Immunocytochemistry in Xenopus. Michael W. Klymkowsky. 2018. [45]
- Microinjection of mRNAs and Oligonucleotides. Sally A. Moody. 2018. [46]
- Microinjection of DNA Constructs into Xenopus Embryos for Gene Misexpression and cis-Regulatory Module Analysis. Yuuri Yasuoka and Masanori Taira. 2019. [47]
- Whole-Mount Immunofluorescence for Visualizing Endogenous Protein and Injected RNA in Xenopus Oocytes. Samantha P. Jeschonek and Kimberly L. Mowry. 2018. [48]
- Isolation of Xenopus Oocytes. Karen Newman, Tristan Aguero, and Mary Lou King. 2018. [49]
- Microinjection of Xenopus Oocytes. Tristan Aguero, Karen Newman, and Mary Lou King. 2018. [50]
- Oocyte Host-Transfer and Maternal mRNA Depletion Experiments in Xenopus. Douglas W. Houston. 2018. [51].
- Applying Tensile and Compressive Force to Xenopus Animal Cap Tissue. Georgina K. Goddard, Nawseen Tarannum, and Sarah Woolner. 2020. [52].
- RNAi-Mediated Loss of Function of Xenopus Immune Genes by Transgenesis. Eva-Stina Edholm and Jacques Robert. 2018. [53]
- Flow Cytometric Analysis of Xenopus Immune Cells. Eva-Stina Edholm. [54]
- Isolation and Demembranation of Xenopus Sperm Nuclei. James W. Hazel and Jesse C. Gatlin. 2018. [55]
- Patch-Clamp and Perfusion Techniques to Study Ion Channels Expressed in Xenopus Oocytes. Guohui Zhang and Jianmin Cui. 2018. [56]
- Heterologous Protein Expression in the Xenopus Oocyte. Jonathan S. Marchant. 2018. [57]
- Isolation and Analysis of Xenopus Germinal Vesicles. Garry T. Morgan. 2018. [58]
- Xenopus Tadpole Tissue Harvest. Matthew D. Patmann, Leena H. Shewade, Katelin A. Schneider, and Daniel R. Buchholz. 2017 [59].
- In Vivo Transfection of Naked DNA into Xenopus Tadpole Tail Muscle. Lindsey Marshall, Fabrice Girardot, Barbara A. Demeneix, and Laurent Coen. 2017. [60]
- Cell Proliferation Analysis during Xenopus Metamorphosis: Using 5-Ethynyl-2-Deoxyuridine (EdU) to Stain Proliferating Intestinal Cells. Morihiro Okada and Yun-Bo Shi. 2017. [61]
'Omics
- Transcriptomics and Proteomics Methods for Xenopus Embryos and Tissues. Michael J. Gilchrist, Gert Jan C. Veenstra, and Ken W.Y. Cho. 2020. ''Topic Introduction'' [62]
- Mass Spectrometry-Based Absolute Quantification of Single Xenopus Embryo Proteomes. Rik G.H. Lindeboom, Arne H. Smits, Matteo Perino, Gert Jan C. Veenstra, and Michiel Vermeulen. [63]
- INTACT Proteomics in Xenopus. Lauren Wasson, Nirav M. Amin, and Frank L. Conlon. 2019. [64]
Genomes, Chromosomes, DNA, Chromatin and Epigenetics
- In Vitro Transcription Systems. Michael R. Green and Joseph Sambrook. [65]
- ChIP-Sequencing in Xenopus Embryos. Saartje Hontelez, Ila van Kruijsbergen, and Gert Jan C. Veenstra. 2019. [66]
- Chromatin Characterization in Xenopus laevis Cell-Free Egg Extracts and Embryos. Wei-Lin Wang, Takashi Onikubo, and David Shechter. 2019. [67].
- Analysis of Chromatin Binding of Ectopically Expressed Proteins in Early Xenopus Embryos. Laura J.A. Hardwick and Anna Philpott. 2019. [68].
- Analysis of Phosphorylation Status of Ectopically Expressed Proteins in Early Xenopus Embryos. Laura J.A. Hardwick and Anna Philpott. 2019. [69].
- Assessing Ubiquitylation of Individual Proteins Using Xenopus Extract Systems. Gary S. McDowell and Anna Philpott. 2019. [70]
- Generating a Three-Dimensional Genome from Xenopus with Hi-C. Ian K. Quigley and Sven Heinz. 2019. [71]
- An RNA-Seq Protocol for Differential Expression Analysis. Nick D.L. Owens, Elena De Domenico, and Michael J. Gilchrist. 2019. [72]
- DNase-seq to Study Chromatin Accessibility in Early Xenopus tropicalis Embryos. Jin Sun Cho, Ira L. Blitz, and Ken W.Y. Cho. 2019 [73]
- Mapping Chromatin Features of Xenopus Embryos. George E. Gentsch and James C. Smith. 2019. [74]
- Reconstituting Nuclear and Chromosome Dynamics Using Xenopus Extracts. Susannah Rankin. 2019 [75]
- Extracts for Analysis of DNA Replication in a Nucleus-Free System. Justin Sparks and Johannes C. Walter. 2019. [76]
- Endoplasmic Reticulum Network Formation with Xenopus Egg Extracts. Songyu Wang, Fabian B. Romano, and Tom A. Rapoport. 2019. [77].
- Chromosome Cohesion and Condensation in Xenopus Egg Extracts. Eulália M.L. da Silva and Susannah Rankin. 2019. [78]
- Chromatin Interaction Analysis Using Paired-End-Tag (ChIA-PET) Sequencing in Tadpole Tissues. Nicolas Buisine, Xiaoan Ruan, Yijun Ruan, and Laurent M. Sachs. 2018. [79]
- Chromatin Immunoprecipitation for Chromatin Interaction Analysis Using Paired-End-Tag (ChIA-PET) Sequencing in Tadpole Tissues. 2018. Nicolas Buisine, Xiaoan Ruan, Yijun Ruan, and Laurent M. Sachs. [80]
Endocrinology, Toxicology and Metamorphosis
- Methods for Investigating the Larval Period and Metamorphosis in Xenopus. Daniel R. Buchholz and Yun-Bo Shi. ''Topic Introduction'' [81].
- Frog Embryo Teratogenesis Assay—Xenopus (FETAX): Use in Alternative Preclinical Safety Assessment. Douglas J. Fort and Michael Mathis. 2018. [82]
- Following Endocrine-Disrupting Effects on Gene Expression in Xenopus laevis. Petra Spirhanzlova, Michelle Leemans, Barbara A. Demeneix, and Jean-Baptiste Fini. 2019. Full-text link: [83].
- Larval Thymectomy of Xenopus laevis. Sara Mashoof, Breanna Breaux, and Michael F. Criscitiello. 2018. [84]
Cell-free extracts, Cell-free systems
- The Use of Cell-Free Xenopus Extracts to Investigate Cytoplasmic Events. Romain Gibeaux and Rebecca Heald. 2019.''Topic Introduction'' [85]
- Calculating the Degradation Rate of Individual Proteins Using Xenopus Extract Systems. Gary S. McDowell and Anna Philpott. 2019. [86].
- Special Considerations for Making Explants and Transplants with Xenopus tropicalis. 2019. Marilyn Fisher and Robert M. Grainger. [87]. Supplemental Material [88].
- Filopodia-Like Structure Formation from Xenopus Egg Extracts. Helen M. Fox and Jennifer L. Gallop. 2019. [89].
- Centromere and Kinetochore Assembly in Xenopus laevis Egg Extract. Julio C. Flores Servin and Aaron F. Straight. 2018. [90]
- Dissecting Protein Complexes in Branching Microtubule Nucleation Using Meiotic Xenopus Egg Extracts. Jae-Geun Song and Sabine Petry. 2018. [91]
- Protein Immunodepletion and Complementation in Xenopus laevis Egg Extracts. Christopher Jenness, David J. Wynne, and Hironori Funabiki. 2018. [92]
- Analysis of Mitotic Checkpoint Function in Xenopus Egg Extracts. Yinghui Mao. 2018 [93]
- Chemical Screening Using Cell-Free Xenopus Egg Extract. Matthew R. Broadus and Ethan Lee. 2018. [94]
- Robustly Cycling Xenopus laevis Cell-Free Extracts in Teflon Chambers. Jeremy B. Chang and James E. Ferrell Jr. [95]
- Microfluidic Encapsulation of Demembranated Sperm Nuclei in Xenopus Egg Extracts. John Oakey and Jesse C. Gatlin. 2018. [96]
- Nucleus Assembly and Import in Xenopus laevis Egg Extract. Pan Chen and Daniel L. Levy. 2018. [97]
- Assembly of Spindles and Asters in Xenopus Egg Extracts. Christine M. Field and Timothy J. Mitchison. 2018. [98]
- Preparation of Cellular Extracts from Xenopus Eggs and Embryos. Matthew C. Good and Rebecca Heald. 2018. [99]
Immunobiology
- Raising Antibodies for Use in Xenopus. Maya Z. Piccinni and Matthew J. Guille. 2020. [100]
- Purifying Antibodies Raised against Xenopus Peptides. Maya Z. Piccinni and Matthew J. Guille. 2020. [101]
- Assessing the Immune Response When Raising Antibodies for Use in Xenopus. Maya Z. Piccinni and Matthew J. Guille. 2020. [102].
- Lymphocyte Deficiency Induced by Sublethal Irradiation in Xenopus. Louise A. Rollins-Smith and Jacques Robert. 2019. [103].
- Skin Grafting in Xenopus laevis: A Technique for Assessing Development and Immunological Disparity. Yumi Izutsu. 2019. [104]
- Adoptive Transfer of Fluorescently Labeled Immune Cells in Xenopus. Kun Hyoe Rhoo and Jacques Robert. [105]
- Assessing Antibody Responses to Pathogens or Model Antigens in Xenopus by Enzyme-Linked Immunosorbent Assay (ELISA). Francisco De Jesús Andino and Jacques Robert. 2019 [106]
- In Vivo Assessment of Neural Precursor Cell Cycle Kinetics in the Amphibian Retina. Morgane Locker and Muriel Perron2019. [107], Supplemental Material [108]
- Protein Immunodepletion and Complementation in Xenopus laevis Egg Extracts. Christopher Jenness, David J. Wynne, and Hironori Funabiki. 2018. [109]
- Elicitation of Xenopus laevis Tadpole and Adult Frog Peritoneal Leukocytes. Leon Grayfer. 2018. [110]
- RNAi-Mediated Loss of Function of Xenopus Immune Genes by Transgenesis. Eva-Stina Edholm and Jacques Robert. 2018. [111]
- Flow Cytometric Analysis of Xenopus Immune Cells. Eva-Stina Edholm. [112]
Neurobiology and tissue regeneration
- Ex Vivo Eye Tissue Culture Methods for Xenopus. Jonathan J. Henry, Kimberly J. Perry, and Paul W. Hamilton. 2019. [113].
- Methods for Examining Lens Regeneration in Xenopus. Jonathan J. Henry, Kimberly J. Perry, and Paul W. Hamilton. 2019. [114]
- Tracing Central Nervous System Axon Regeneration in Xenopus. Kurt M. Gibbs and Ben G. Szaro. 2018. [115]
- Cell Transplantation as a Method to Investigate Spinal Cord Regeneration in Regenerative and Nonregenerative Xenopus Stages. Emilio E. Méndez-Olivos and Juan Larraín. 2018. [116]
- Infrared Laser-Mediated Gene Induction at the Single-Cell Level in the Regenerating Tail of Xenopus laevis Tadpoles. Riho Hasugata, Shinichi Hayashi, Aiko Kawasumi-Kita, Joe Sakamoto, Yasuhiro Kamei, and Hitoshi Yokoyama. 2018. [117]
- Rod-Specific Ablation Using the Nitroreductase/Metronidazole System to Investigate Regeneration in Xenopus. Reyna I. Martinez-De Luna and Michael E. Zuber. 2018. [118]
- Studies of Limb Regeneration in Larval Xenopus. Anthony L. Mescher and Anton W. Neff. 2019. [119].
- Bulk Electroporation-Mediated Gene Transfer into Xenopus Tadpole Brain. Cristina Sáenz de Miera, Ethan Parr, and Robert J. Denver. 2018. [120] . Supplemental Material [121].
- nverse Drug Screening of Bioelectric Signaling and Neurotransmitter Roles: Illustrated Using a Xenopus Tail Regeneration Assay. Kelly G. Sullivan and Michael Levin. 2018. [122]
Fate mapping, Explants and Transplants
- Dissecting and Culturing Animal Cap Explants. Kevin S. Dingwell and James C. Smith. 2018. [123]
- Cranial Neural Crest Explants. Hélène Cousin and Dominique Alfandari. 2018. [124]
- Einsteck Transplants. Hélène Cousin. 2019 [125]
- Cleavage Blastomere Deletion and Transplantation to Test Cell Fate Commitment in Xenopus. Sally A. Moody. 2019. [126]
- Cleavage Blastomere Explant Culture in Xenopus. Sally A. Moody. 2019. [127]
- Lineage Tracing and Fate Mapping in Xenopus Embryos. Sally A. Moody. 2018. [128]
- Collagen-Embedded Tumor Transplantations in Xenopus laevis Tadpoles. Maureen Banach and Jacques Robert. 2017 [129]
- Organ Culture of the Xenopus Tadpole Intestine. Atsuko Ishizuya-Oka. 2017 [130]
CSHL Recipes
- Embryo Lysis Buffer (Xenopus). (Recipe 1) CSHLP. 2019 [131]
- Xenopus Embryo Lysis Buffer. (Recipe 2). CSHLP. 2019 [132]
- Lysis Buffer for Xenopus (recipe3) CSHLP. 2019. [133]
- DNA Isolation Buffer. CSHLP. 2019 [134]
- Lysis Buffer for Xenopus Hi-C. CSHLP 2019. [135]
- RIPA Buffer for Xenopus. CSHLP 2019. [136]
- Amphibian Serum-Free (ASF) Medium Supplemented with Fetal Bovine Serum (FBS). CSHL 2019. [137]
- Xenopus Eye Culture Medium. CSHLP 2019. [138]
- Marc's Modified Ringer's (MMR) for Xenopus (1×). CSHLP. 2018. [139]
- Marc's Modified Ringer's (MMR) for Xenopus (20×). CSHLP. 2019. [140]
- Injection Buffer for Xenopus. CSHLP 2019. [141]
- TE Buffer for Xenopus. CSHLP 2019. [142]
- Proteinase K Buffer for Xenopus Oocytes. CSHLP 2018. [143]
- PBT/PBT-Plus for Xenopus Oocytes. CSHL. 2018. [144]
- Energy Mix for Xenopus Egg Extracts. CSHL 2018. [145]
- Xenopus Oocyte Culture Medium (XOCM).CSHL 2018.[146]
- PBT for Xenopus Oocyte FISH.CSHL 2018. [147]
- Blocking Solution for Xenopus Oocytes. CSHL 2018. [148]
- Aldehyde Fixative (MEMFA). CSHL 2018. [149]
- Staining Buffer for Xenopus Embryos. CSHL 2017. [150]
- Blocking Solution for Xenopus Embryos. CSHL 2017. [151]
- Embryonic Xenopus Culture Media (CM). CSHL 2017. [152]
- Hybridization Buffer (HB) for Xenopus Embryos. CSHL 2017. [153]
Cold Spring Harbor Xenopus Protocols 2007. Edited by Hazel Sive
- Housing and Feeding of Xenopus laevis - Sive et al. [154]
- Inducing Ovulation in Xenopus laevis - Sive et al. [155]
- Xenopus laevis In Vitro Fertilization and Natural Mating Methods - Sive et al. [156]
- Egg Collection and In Vitro Fertilization of the Western Clawed Frog Xenopus tropicalis - Showell & Conlon [157]
- Isolation of Xenopus Oocytes - Sive et al. [158]
- Isolating Xenopus laevis Testes - Sive et al. [159]
- Dejellying Xenopus laevis Embryos - Sive et al. [160]
- Removing the Vitelline Membrane from Xenopus laevis Embryos - Sive et al. [161]
- Microinjection of Xenopus Embryos - Sive et al. [162]
- Defolliculation of Xenopus Oocytes - Sive et al. [163]
- Microinjection of Xenopus Oocytes - Sive et al. [164]
- Animal Cap Isolation from Xenopus laevis - Sive et al. [165]
- Xenopus laevis Keller Explants - Sive et al. [166]
- Microinjection of RNA and Preparation of Secreted Proteins from Xenopus Oocytes - Sive et al. [167]
- Calibration of the Injection Volume for Microinjection of Xenopus Oocytes and Embryos - Sive et al. [168]
- Isolation of DNA from Red Blood Cells in Xenopus - Sive et al. [169]
- Investigating Morphogenesis in Xenopus Embryos: Imaging Strategies, Processing, and Analysis - Kim & Davidson [170]
- Low-Magnification Live Imaging of Xenopus Embryos for Cell and Developmental Biology - Wallingford [171]
- High-Magnification In Vivo Imaging of Xenopus Embryos for Cell and Developmental Biology - Keiserman et al. [172]
- Preparation of Fixed Xenopus Embryos for Confocal Imaging - Wallingford [173]
- Whole-Mount Fluorescence Immunocytochemistry on Xenopus Embryos - Lee et al. [174]
- Generation of Transgenic Xenopus laevis - Kroll & Amaya - [175] [176] [177]
- In Vivo Time-Lapse Imaging of Neuronal Development in Xenopus - Ruthazer et al. [178]
- Photoconversion for Tracking the Dynamics of Cell Movement in Xenopus laevis Embryos - Chernet et al. [179]
- Single-Cell Electroporation in Xenopus - Liu & Haas [180]
- Imaging Axon Pathfinding in Xenopus In Vivo - Leung & Holt [181]
- A Versatile Protocol for mRNA Electroporation of Xenopus laevis Embryos - Chernet & Levin [182]
General Research Protocols
Animal Husbandry
- obtain frogs and frog housing equipment [183]
- Xenopus Express links [184] - Xenopus Express husbandry [185]
- 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). [186]
- A method for generating transgenic frog embryos. Ishibashi S, Kroll KL, Amaya E. Methods Mol Biol. 2008;461:447-66. [187]
- A method for generating transgenic frog embryos. Amaya E, Kroll KL. Methods Mol Biol. 1999;97:393-414. [188]
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. [189] [190]
- Akkers RC, Jacobi UG, Veenstra GJ.
- Chromatin immunoprecipitation in early Xenopus laevis embryos., Dev Dyn. 2009 Jun;238(6):1422-32. [191] [192]
- 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)