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RNA-RNA In Situ Hybridization Using DIG-Labeled Probes Protocol - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_172-177.pdf

Category: Cytogenetics Protocols: RNA In Situ Hybridization Protocols

RNA-RNA in situ hybridization using DIG-labeled probes: the effect of high molecular weight polyvinyl alcohol on the alkaline phosphatase indoxyl-nitroblue tetrazolium reaction. RNA-RNA in situ hybridization protocol using alkaline phosphatase-conjugated digoxigenin-(DIG-) labeled probes is presented. The addition of polyvinyl alcohol (PVA) of high molecular weight (40 – 100 kD) to the BCIP-NBT detection system enhances the alkaline phosphatase reaction and prevents... - [Read RNA-RNA In Situ Hybridization Using DIG-Labeled Probes Protocol]

Single Colour Fish Analysis Method - http://www.sanger.ac.uk/HGP/methods/cytogenetics/single_colour.shtml

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Single colour detection for biotinylated probes. Sanger Institute - [Read Single Colour Fish Analysis Method]

Southern Hybridization of Radiolabeled Probes to Nucleic Acids Immobilized on Membranes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4044

Category: Nucleic Acid Detection Protocols: Nucleic Acid Blotting Protocols

Protocol for southern hybridization of radiolabeled probes to nucleic acids immobilized on membranes. Protocol describes how to carry out Southern hybridizations at high stringency in phosphate-SDS buffers. Although a wide variety of formats are available, most Southern hybridizations are carried out in heat-sealable bags, roller bottles, or plastic boxes. - [Read Southern Hybridization of Radiolabeled Probes to Nucleic Acids Immobilized on Membranes Protocol]

Synthesis of cDNA Probes from mRNA Using Random Oligonucleotide Primers Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3868

Category: DNA Protocols: cDNA Library Protocols

Protocol describes the generation of cDNA probes from poly(A)+ mRNA using random oligonucleotide primers. Probes of this type are used for differential screening of cDNA libraries. - [Read Synthesis of cDNA Probes from mRNA Using Random Oligonucleotide Primers Protocol]

Synthesis of Radiolabeled, Subtracted cDNA Probes Using Oligo(dT) as a Primer Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3869

Category: PCR Protocols: cDNA Synthesis Protocols

Protocol describes the preparation of subtracted cDNA probes by hybridization to an mRNA driver, followed by purification of the single-stranded radiolabeled cDNA by hydroxyapatite chromatography. Before preparing the probe, it is a good idea to have filters (which contain the cDNA library to be screened) ready to hybridize. - [Read Synthesis of Radiolabeled, Subtracted cDNA Probes Using Oligo(dT) as a Primer Protocol]

Synthesis of Single-stranded DNA Probes of Defined Length from Bacteriophage M13 Templates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4061

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

A synthetic oligonucleotide annealed to single-stranded DNA derived from a recombinant bacteriophage M13 or phagemid template is used to prime the synthesis of complementary radiolabeled DNA. Synthesis is catalyzed by the Klenow fragment of E. coli DNA polymerase I, which extends the annealed primer for various distances along the single-stranded template DNA. - [Read Synthesis of Single-stranded DNA Probes of Defined Length from Bacteriophage M13 Templates Protocol]

Synthesis of Single-stranded DNA Probes of Heterogeneous Length from Bacteriophage M13 Templates - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3546

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Technique yields a heterogeneous population of short radiolabeled molecules 200-300 nucleotides in length. These probes are synthesized, as in Synthesis of Single-stranded DNA Probes of Defined Length from Bacteriophage M13 Templates, by extension of an oligonucleotide primer on a single-stranded DNA template. The radiolabeled products of the reaction are then separated from the template by electrophoresis through a denaturing gel from which they are eluted directly into hybridization buffer. - [Read Synthesis of Single-stranded DNA Probes of Heterogeneous Length from Bacteriophage M13 Templates]

Time-Lapse Cinematography in Living Drosophila Tissues: Preparation of Material - http://www.cshprotocols.org/cgi/content/abstract/2006/29/pdb.prot4602

Category: Microscopy Protocols: In Vivo Imaging Protocols

Sophisticated fluorescence microscopy methods & equipment, now allow cellular events to be studied at high resolution in living material. The studying of living fly tissues presents unique difficulties in keeping the cells alive, introducing fluorescent probes, & imaging through thick hazy cytoplasm. This protocol outlines the preparation of major tissue types amenable to study by time-lapse cinematography and different methods for keeping them alive. - [Read Time-Lapse Cinematography in Living Drosophila Tissues: Preparation of Material]

Two-colour detection for combinations of biotinylated and digoxigenin-labelled probes - http://www.sanger.ac.uk/HGP/methods/cytogenetics/two_colour.shtml

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Two-colour detection for combinations of biotinylated and digoxigenin-labelled probes. Sanger Institute - [Read Two-colour detection for combinations of biotinylated and digoxigenin-labelled probes]

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Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

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