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Detection of Even-Skipped Transcripts in Drosophila Embryos with PCR/DIG-Labeled DNA Probes Protocol - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_216-219.pdf

Category: Cytogenetics Protocols: In Situ Hybridization Tissue Sections Protocols

Protocol for detection of even-skipped transcripts in drosophila embryos with PCR/DIG-labeled DNA probes. This protocol has been used to detect the transcript distribution of a number of genes by in situ hybridization, including evenskipped and seven-up, in whole mount Drosophila embryos, and engrailed Antennapedia in whole mount grasshopper embryos. Includes: Probe labeling; Evaluation of labeling reaction; Preparation of embryos, hybridization and detection. - [Read Detection of Even-Skipped Transcripts in Drosophila Embryos with PCR/DIG-Labeled DNA Probes Protocol]

ES cell culture protocols (2005 update) PDF Excellent - http://www.mshri.on.ca/cores/ESfacility/ESculture.pdf

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: ES Cell Growth Media and Requirements

ES cell culture protocols (2005 update) PDF Excellent. ES cell facility, Samuel Lunenfeld Research Institute, Mount Sinai Hospital. - [Read ES cell culture protocols (2005 update) PDF Excellent]

Mount Sections in Aqueous Mountant Protocol - http://www.nottingham.ac.uk/pathology/protocols/mountaqu.html

Category: Immunohistochemistry Protocols: Coated Treated Slides Protocols

Protocol for mount sections in aqueous mountant. - [Read Mount Sections in Aqueous Mountant Protocol]

Mount Sections in DPX Protocol - http://www.nottingham.ac.uk/pathology/protocols/mountdpx.html

Category: Immunohistochemistry Protocols: Coated Treated Slides Protocols

Protocol for mount sections in DPX. - [Read Mount Sections in DPX Protocol]

Preparing Early Whole-Mount Drosophila Embryos for Immunostaining Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4524

Category: Drosophila Protocols: Drosophila Staining Protocols

Early embryos (0-17 hours or until cuticle formation) are treated with a mixture of organic solvents, formaldehyde, and alcohols, as described here. The cuticles of late-stage embryos are usually opened by sonication. Tissues from more advanced stages of development are normally dissected by hand and then fixed and stained in a standard paraformaldehyde/detergent combination - [Read Preparing Early Whole-Mount Drosophila Embryos for Immunostaining Protocol]

Preparing Late Whole-Mount Drosophila Embryos for Immunostaining Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4525

Category: Drosophila Protocols: Drosophila Staining Protocols

Early and late embryos are treated with a mixture of organic solvents, formaldehyde, and alcohols. The cuticles of late-stage embryos (17-22 hours or until hatching) are usually opened by sonication, as described here. Tissues from later stages of development are normally dissected by hand and then fixed and stained in a standard paraformaldehyde/detergent combination. - [Read Preparing Late Whole-Mount Drosophila Embryos for Immunostaining Protocol]

Protocol for Nuclear Staining of Plants for Confocal Microscopy - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4685

Category: Microscopy Protocols: Confocal Microscopy Protocols

Protocol describes a useful way to observe the development of embryos, as well as meristems & young primordia developing at the shoot apex by confocal microscopy after staining the nuclei with propidium iodide. The number of cells can be exactly quantified in a meristem or in young primordia. Because embryonic & meristematic cells are largely filled out by their nuclei, it is easier to image only the nuclei. This method allows analysis of whole-mount material, which is more easily reconstructed. - [Read Protocol for Nuclear Staining of Plants for Confocal Microscopy]

RNA In Situ Hybridization of Dissected Gonads with Digoxygenin-Labeled Probes Protocol - http://www.genetics.wustl.edu/tslab/Protocols/gonad_in_situ.html

Category: C. elegans Protocols

Protocol for RNA in situ hybridization of dissected gonads with digoxygenin-labeled probes. Includes: Preparation of single-stranded probes from cloned cDNA; Dissections and Fixation; Proteinase K digestion; Hybridization; Making a Mount. - [Read RNA In Situ Hybridization of Dissected Gonads with Digoxygenin-Labeled Probes Protocol]

RNA Whole Mount In Situ Hybridization Protocol - http://genetics.med.harvard.edu/~cepko/protocol/ctlab/ish.ct.htm

Category: Cytogenetics Protocols: In Situ Hybridization Tissue Sections Protocols

Protocol for RNA whole mount in situ hybridization. Includes: Embryo preparation; Prehybridization and Hybridization; Post-hybridization washes, blocking, and antibody incubation; Post-antibody washes; Color development. - [Read RNA Whole Mount In Situ Hybridization Protocol]

Staining of Tissue Slices for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4507

Category: RNAi Technology Protocols: RNAi Avian Embryos Protocols

Protocol describes a method to stain nerve fibers in tissue slices of avian embryos using an antibody against the 160-kD subunit of neurofilaments. This allows the comparison of the branching pattern of motor and sensory neurons between control and experimental embryos. The tissue is cut in slices using a vibratome or tissue slicer. The protocol is suitable for older embryos after approximately stage 33 and regions that are not accessible by whole-mount analysis. - [Read Staining of Tissue Slices for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos]

Staining of Tissue Slices for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4507

Category: Avian Bird Protocols

Transcript In Situ Hybridization of Whole-Mount Embryos for Phenotype Analysis of RNAi-Treated - http://www.cshprotocols.org/cgi/content/full/2006/21/pdb.prot4519

Category: Cytogenetics Protocols: In Situ Hybridization Protocols

This protocol fixes and prepares embryos for in situ hybridization to visualize transcript expression patterns. It is a modification of the method developed by Tautz and Pfeifle for whole-mount in situ analysis of embryos. Use of the standard hybridization protocol on RNAi-treated embryos results in high background staining, which makes visualization of transcript expression patterns practically impossible. The following modifications eliminate this problem and allow visualization of transcript. - [Read Transcript In Situ Hybridization of Whole-Mount Embryos for Phenotype Analysis of RNAi-Treated]

Whole Mount Fluorescence in Situ Hybridization (FISH) of Repetitive DNA Sequences on Interphase - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_220-228.pdf

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol for whole mount fluorescence in situ hybridization (FISH) of repetitive DNA sequences on interphase nuclei of the small cruciferous plant Arabidopsis thaliana. Includes: Seed sterilization and germination; Tissue fixation; Labeling of the probe DNA; Pretreatment; In situ hybridization; Pre-absorption of antibodies; Posthybridization washes; Immunocytochemical detection; Direct detection; Indirect detection; Staining and mounting; Fluorescence microscopy. - [Read Whole Mount Fluorescence in Situ Hybridization (FISH) of Repetitive DNA Sequences on Interphase]

Whole Mount Fluorescent Antibody Staining of Drosophila Embryos Protocol - http://vosshall.rockefeller.edu/protocols/EmbryoImmunofluo.pdf

Category: Drosophila Protocols: Drosophila Staining Protocols

Protocol for whole mount fluorescent antibody staining of Drosophila embryos. - [Read Whole Mount Fluorescent Antibody Staining of Drosophila Embryos Protocol]

Whole Mount Fluorescent Antibody Staining of Drosophila Larvae Protocol - http://vosshall.rockefeller.edu/protocols/LarvaFlu.pdf

Category: Drosophila Protocols: Drosophila Staining Protocols

Protocol for whole mount fluorescent antibody staining of Drosophila larvae. - [Read Whole Mount Fluorescent Antibody Staining of Drosophila Larvae Protocol]

Whole Mount In Situ Hybridization for the Detection of mRNA in Drosophila Embryos Protocol - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_208-215.pdf

Category: Drosophila Protocols: Drosophila Genetics Protocols

Protocol for whole mount in situ hybridization for the detection of mRNA in Drosophila embryos. Includes: Embryo collection and fixation; Prehybridization treatment; Hybridization; Washing and detection. - [Read Whole Mount In Situ Hybridization for the Detection of mRNA in Drosophila Embryos Protocol]

Whole Mount In Situ Hybridization of Mouse Protocol - http://www.hhmi.ucla.edu/derobertis/protocol_page/in_situ.html

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol for whole mount in situ hybridization of mouse. Incluses: Preparation of the probe; General Comments; Preparation of embryos; In Situ hybridization. - [Read Whole Mount In Situ Hybridization of Mouse Protocol]

Whole-Mount DAPI Staining and Measurement of DNA Content in Plant Cells - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4684

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

During development many plant cells undergo endoreduplication, whereby ploidy increases to a multiple of the normal 2C content. For eg., trichome development is accompanied by an increase in ploidy to 32C, indicating that trichome cells undergo four rounds of endoreduplication. Protocol describes DNA levels, and hence developmental progress in the corresponding cells, are measured by staining the DNA with a fluorescent marker and then quantifying the fluorescence of individual nuclei. - [Read Whole-Mount DAPI Staining and Measurement of DNA Content in Plant Cells]

Whole-Mount In Situ Hybridization for the Detection of RNA in C. elegans Embryos Protocol - http://cobweb.dartmouth.edu/~ambros/worms/14.html

Category: C. elegans Protocols

Protocol for whole-mount in situ hybridization for the detection of RNA in C. elegans embryos. Includes: Collection of Embryos; Permeabilization and fixation of embryos; Prehybridization; Hybridization; Post-hybridization washes; Probe Detection; Double-labeling; Interpretation. - [Read Whole-Mount In Situ Hybridization for the Detection of RNA in C. elegans Embryos Protocol]

Whole-Mount Preparations for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4506

Category: RNAi Technology Protocols: RNAi Avian Embryos Protocols

Protocol describes a method for staining nerve fibers in whole-mount preparations of avian embryos using an antibody against the 160-kD subunit of neurofilaments. This allows the comparison of the branching pattern of motor and sensory neurons between control and experimental embryos. This protocol has been successfully applied for embryos at different stages up to about stage 33 (7 days of incubation). - [Read Whole-Mount Preparations for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos]

Whole-Mount Preparations for Analysis of Axonal Pathfinding in dsRNA-Treated Avian Embryos Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4506

Category: Avian Bird Protocols

Xenopus Whole Mount In Situ Hybridization Protocols - http://tropicalis.berkeley.edu/home/gene_expression/in-situ/insitu.html

Category: Xenopus Protocols: Xenopus Genetics Protocols

Protocols for Xenopus whole mount in situ hybridization. Includes: Protocols for Making Antisense RNA probe (dig labelled); Protocols for In Situ Hyb. - [Read Xenopus Whole Mount In Situ Hybridization Protocols]

Articles (1-10 of 12)

Populational Analysis of Genomic DNA Protocol

A protocol on the populational Analysis of genomic DNA.

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

Constructing Random-Peptide Libraries for Phage Display

The protocol describes the construction of a large (10^8 to 10^10) primary random peptide phage library in fUSE5 or f88-4 strains.

DNA Ligation Protocol

The DNA Ligation protocol described here contains the steps required to join together using ligase enzyme both plasmid DNA and insert DNA fragments in order to create a new plasmid. This new ligated plasmid can be transformed after into competent bacteria to produce DNA for mini, midi or maxi-prep isolation.