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Category: Main/IN SITU HYBRIDIZATION/Protocols


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35S-labeled probe  Popular
Description: In Situ Hybridization using 35S-labeled probe, Detection by emulsion or film autoradiography. Frozen Sections.
Added on: 23-Nov-2004 Hits: 5215 Rating: 10 (1 Vote)
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ACCUMULATION & FIXATION OF PLANT METAPHASE CHROMOSOMES 
Description: ACCUMULATION & FIXATION OF PLANT METAPHASE CHROMOSOMES
Added on: 17-Dec-2009 Hits: 371
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Anderson Lab In Situ Hybridization Protocols  Popular
Description: In Situ Hybridization of Frozen Sections, Whole Mount In Situ Hybridization, In Situ Hybridization on Cultured Cells and Additional Techniques
Added on: 07-Oct-2004 Hits: 13614 Rating: 6 (2 Votes)
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Autoradiography for mRNA detection in mouse embryo tissue sections 
Description: Protocols from Oxford Practical Approach Series
Added on: 07-Oct-2004 Hits: 2939
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BUFFERS AND OTHER SOLUTIONS 
Description: BUFFERS AND OTHER SOLUTIONS
Added on: 17-Dec-2009 Hits: 668
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CHECKING BIOTIN AND DIGOXIGENIN LABELED PROBE  
Description: CHECKING BIOTIN AND DIGOXIGENIN LABELED PROBE
Added on: 17-Dec-2009 Hits: 597
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Chick, mouse, and Xenopus two colour whole mount ISH 
Description: Protocols from Oxford Practical Approach Series. Chick, mouse, and Xenopus two colour whole mount in situ hybridization staining with Fast Red and TrueBlue
Added on: 07-Oct-2004 Hits: 3442 Rating: 1 (1 Vote)
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Chromosome In Situ Hybridization using biotin labeled probes 
Description: Chromosome In Situ Hybridization using biotin labeled probes. Preparation of Chromosome Squashes
Added on: 01-Dec-2004 Hits: 4517
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Chromosome preparation for ISH 
Description: Chromosome preparation for in situ hybridization (ISH)
Added on: 07-Oct-2004 Hits: 2814
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Culture and harvesting of human metaphase chromosomes 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 2063
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DENATURATION AND HYBRIDIZATION  
Description: DENATURATION AND HYBRIDIZATION
Added on: 17-Dec-2009 Hits: 555
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Denaturation of DNA in ultrathin sections of Lowicryl K4M embedded material 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 1580
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Detection and amplification of FISH signal 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 4621
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Detection of mRNA by in situ hybridization  Popular
Description: The following page describes the current favorite non-radioactive in situ hybridization technique used by Dr. G.V. Childs at the University of Arkansas for Medical Sciences to detect mRNAs in whole pituitary cells grown in culture.
Added on: 07-Oct-2004 Hits: 12200 Rating: 10 (2 Votes)
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Detection of multiply spliced HIV-1 tat mRNA by RT-PCR in situ hybridization 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 877
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Detection of single-copy HIV-1 DNA by PCR in situ hybridization 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 892
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Detection of single-copy HIV-1 DNA using Real-Time (Taqman) PCR in situ 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 801
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DIG-labeled probe 
Description: In Situ Hybridization using DIG-labeled probe, Detection by AP (alkaline phosphatase), FITC or rhodamine. Paraffin Sections.
Added on: 23-Nov-2004 Hits: 4627 Rating: 10 (2 Votes)
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DNA: DNA in situ hybridization 
Description: Procedures for in situ hybridization of labeled DNA probes to spread or sectioned chromosomes and nuclei (DNA) are described here. Double-target experiments are recommended for most applications, while multiple-target or reprobing of preparations are possible if more information is required.
Added on: 17-Dec-2009 Hits: 498
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ETHANOL PRECIPITATION  
Description: ETHANOL PRECIPITATION
Added on: 17-Dec-2009 Hits: 453
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FISH Guide and Troubleshooting 
Description: FISH Guide and Troubleshooting
Added on: 13-Feb-2009 Hits: 944
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FISH hybridization and subsequent washes 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 2187
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FISH protocols for Drosophila 
Description: Fluorescent in situ hybridization (FISH) is the standard method for visualizing the spatial distribution of RNA. While traditional histochemical RNA detection methods suffered from limitations in resolution or sensitivity, the recent development of peroxidase-mediated tyramide signal amplification (TSA) provides strikingly enhanced sensitivity and subcellular resolution. In this chapter, we describe optimized FISH protocols for Drosophila embryos and tissues utilizing TSA, either for single genes or in a high-throughput format, which greatly increase the sensitivity, consistency, economy and throughput of the procedure. We also describe variations of the method for RNARNA and RNA-protein co-detection.
Added on: 19-Dec-2007 Hits: 717
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Fixation and pre-treatment of embryos for whole mount hybridization 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 1285
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Fixation and sectioning of cryosections for mRNA detetcion 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 2264
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Fixation and storage of mouse embryos for mRNA detection 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 1061
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Fluorescence In Situ Hybridization (FISH) 
Description: Fluorescence In Situ Hybridization (FISH) on human chromosomes and interphase nuclei.
Added on: 07-Oct-2004 Hits: 2786
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FLUORESCENT DETECTION OF HYBRIDIZATION SITES 
Description: FLUORESCENT DETECTION OF HYBRIDIZATION SITES
Added on: 17-Dec-2009 Hits: 403
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Fluorescent in situ Hybridization (FISH) 
Description: Fluorescent in situ Hybridization (FISH) protocol from Sigma-Aldrich
Added on: 30-Nov-2004 Hits: 2558
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Fluorescent in situ hybridization (FISH) 
Description: ULS(TM)-dGreen and ULS(TM)-Rhodamine labeled probes for translocation, locus specific and chromosome copy number detection
Added on: 10-Dec-2004 Hits: 1237
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FLUORESCENT STAINING OF DNA AND MOUNTING 
Description: FLUORESCENT STAINING OF DNA AND MOUNTING
Added on: 17-Dec-2009 Hits: 473
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GeneDetect Laboratory Protocols 
Description: In Situ Hybridization protocols optimized for use with GeneDetect ® products.
Added on: 06-Oct-2004 Hits: 1397
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GreenStar* BIOTIN-labeled probe 
Description: In Situ Hybridization using GreenStar* BIOTIN-labeled probe in paraffin sections. Detection by AP (alkaline phosphatase) or HRP (Horseradish Peroxidase) with or without tyramide signal amplification (TSA).
Added on: 23-Nov-2004 Hits: 1119
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GreenStar* BIOTIN-labeled probe 
Description: In Situ Hybridization using GreenStar* BIOTIN-labeled probe in frozen sections. Detection by AP (alkaline phosphatase) or HRP (Horseradish Peroxidase) with or without tyramide signal amplification (TSA).
Added on: 23-Nov-2004 Hits: 900
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GreenStar* DIG-labeled probe 
Description: In Situ Hybridization using GreenStar* DIG-labeled probe in paraffin sections. Detection by direct fluorescence, AP (alkaline phosphatase) or tyramide signal amplification (TSA).
Added on: 23-Nov-2004 Hits: 1111
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GreenStar* DIG-labeled probe 
Description: In Situ Hybridization using GreenStar* DIG-labeled probe in frozen sections. Detection by direct fluorescence, AP (alkaline phosphatase) or tyramide signal amplification (TSA).
Added on: 23-Nov-2004 Hits: 958
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GreenStar* FITC-labeled probe 
Description: In Situ Hybridization using GreenStar* FITC-labeled probe, Detection by direct fluorescence, AP (alkaline phosphatase) or tyramide signal amplification (TSA). Frozen Sections.
Added on: 23-Nov-2004 Hits: 918
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Hybridization mixture for cloned and genomic probes 
Description: Hybridization mixture for cloned and genomic probes
Added on: 17-Dec-2009 Hits: 337
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Hybridization of mouse embryo sections for mRNA detection 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 1148
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Hybridization of sections, washing, and detection of probe 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 2403
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Hybridization solution: composition, heat treatment prior to post-embedding ish 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 1336
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Immunolocalization by avidin biotinylated enzyme complex staining of zebrafish embryos after ish 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 955
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Immunolocalization by the peroxidase anti-peroxidase (PAP) method after ish 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 987
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Immunolocalization with a horse-radish peroxidase-conjugated secondary antibody after ish 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 953
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Immunophenotyping simultaneously with RT-PCR in situ hybridization, DNA-PCR ish, and ish 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 850
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In Situ Hybridization 
Description: Collection of ISH/FISH Protocols from Protocol Online.
Added on: 07-Oct-2004 Hits: 3681
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In Situ Hybridization Detection Systems (Vector Laboratories) 
Description: Several different fluorescent or enzyme-based systems are used for detecting labeled nucleic acid probes. These options provide the researcher with flexibility in optimizing experimental systems to achieve highest sensitivity, to avoid potential problems such as endogenous biotin or enzyme activity, or to introduce multiple labels in a single experiment. Such factors as tissue fixation, endogenous biotin or enzyme activity, desired sensitivity, and permanency of record should all be considered when choosing both the optimal probe label and subsequent detection system.
Added on: 07-Oct-2004 Hits: 1377
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In Situ Hybridization Digoxigenin and Fluorecein labeled cRNA Probes 
Description: In Situ Hybridization Digoxigenin and Fluorecein labeled cRNA Probes with double label addendum
Added on: 31-Oct-2004 Hits: 1197
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In situ hybridization of DIG-labelled RNA probes 
Description: Paraffin and Cryo-sections Protocols by Carin Lundmark
Added on: 07-Oct-2004 Hits: 1978
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In situ hybridization of mammalian cells 
Description: In situ hybridization of mammalian cells (RNA and Oligonucleotide probes)
Added on: 07-Oct-2004 Hits: 1415
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In Situ Hybridization of mRNA and Immunofluorescence 
Description: Corbett Lab: In Situ Hybridization of mRNA and Immunofluorescence
Added on: 07-Oct-2004 Hits: 1450
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In situ hybridization of yeast cells 
Description: In situ hybridization of yeast cells (RNA and Oligonucleotide probes)
Added on: 07-Oct-2004 Hits: 819
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In Situ Hybridization on Frozen Sections 
Description: Protocol by Josiah N. Wilcox, Ph.D.
Added on: 07-Oct-2004 Hits: 1401
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In situ Hybridization on imaginal discs 
Description: In situ Hybridization on imaginal discs, with option of histochemical or fluorescence visualization.
Added on: 07-Oct-2004 Hits: 773
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In Situ Hybridization to Tissue Sections 
Description: In Situ Hybridization to Tissue Sections (Kornberg et al., Cell 40:45, 1985)
Added on: 07-Oct-2004 Hits: 1996
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In Situ Hybridization Using Digoxigenin Labeled Probes 
Description: Berkeley Drosophila Genome Project
Added on: 07-Oct-2004 Hits: 1311
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In Situ Hybridization with Nanogold®-Streptavidin 
Description: The small size of Nanogold® probes, their lack of aggregation and their consequent ability to access nuclear antigens makes them an excellent reagent for ultrasensitive in situ hybridization detection of specific DNA sequences both with and without in situ PCR.
Added on: 07-Oct-2004 Hits: 787
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In Situ Hybridization with RNA Probes 
Description: Embryo fixation, Preparing DNA to make RNA probes, Probe synthesis and Hybridization
Added on: 07-Oct-2004 Hits: 1324
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Labelling of probes for FISH by nick translation 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 913
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MITOTIC CHROMOSOME PREPARATIONS 
Description: MITOTIC CHROMOSOME PREPARATIONS FROM MAMMALIAN SUSPENSION CELL CULTURES
Added on: 17-Dec-2009 Hits: 322
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mRNA In Situ Hybridization with Nick-Translated Probes 
Description: mRNA In Situ Hybridization with Nick-Translated Probes (Spector Lab)
Added on: 07-Oct-2004 Hits: 778
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Multiplex Fluorescent mRNA In Situ Hybridization 
Description: Presents information on how to do multiplex fluorescent mRNA in situ hybridization on Drosophila embryos.
Added on: 10-Dec-2004 Hits: 1224 Rating: 10 (1 Vote)
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Nanogold DNA In Situ Hybridization 
Description: This molecular morphological procedure is based on silver/gold-enhanced Nanogold (Nanoprobes, Inc.) and allowed a reliable and very sensitive detection of few copies of human papillomavirus (HPV), cytomegalovirus (CMV), Epstein Barr Virus (EBV), herpes simplex virus (HSV) and other DNA-viruses in routinely formalin-fixed and paraffin-embedded tissues. Other DNA sequences, as well as RNA stainings, have not yet been tested by us. The method described below can be applied as a robust and fast routine method for diagnostic purposes in all cases where "conventional ISH" is not sensitive enough, and where real single-copy sensitivity is not neded. The protocol also works well with Fluoro-Nanogold™ (Nanoprobes, Inc.), thus allowing visualization of the very same preparation by fluorescent microscopy (FISH) and transmitted light microscopy. Nanogold-preparations can be transferred to the electron microscope
Added on: 16-Nov-2004 Hits: 785
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Nonradioactive In Situ Hybridization Application Manual 
Description: This manual concentrates on two labeling systems:
-Indirect methods using digoxigenin (detected by specific antibodies) and biotin (detected by streptavidin)
-Direct methods using fluorescein or other fluorochromes directly coupled to the nucleotide

Added on: 07-Oct-2004 Hits: 1525
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PCR in situ hybridization for a representative tissue control (HLA-DQalpha amplification) 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 824
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Post-embedding in situ hybridization and detection of hybrids 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 767
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Post-hybridization washes after mRNA detection in mouse embryo tissue sections 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 845
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Pre-absorption of antibody 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 993
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Pre-treatment of cryosections for mRNA detection in tissue sections 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 829
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Pre-treatment of sections before mRNA detection 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 875
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Pre-treatment of sections or cells on slides 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 922
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PREPARATION OF ACID CLEANED SLIDES 
Description: PREPARATION OF ACID CLEANED SLIDES
Added on: 17-Dec-2009 Hits: 332
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PREPARATION OF CHROMOSOME SPREADS 
Description: An ideal method of tissue preparation ensures both good specimen morphology and that the target molecules are in the optimum state for probe access and hybridization. DNA:DNA in situ hybridization is usually carried out on chromosome spread preparations where chromosome and nuclei are released from cells and spread on a glass microscope slide. This method yields well separated and enlarged chromosomes with good morphology which can be analyzed in transmitted light or fluorescence microscopes.
Added on: 17-Dec-2009 Hits: 308
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Preparation of Formvar carbon coated grids for ultrathin sections of Lowicryl embedded cells prior t 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 792
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Preparation of labelled whole chromosome paints by DOP-PCR 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 743
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Preparation of lymphoid cells and tissue for molecular and immunological applications 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 715
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Preparation of probes for in situ hybridization 
Description: Preparation of probes for in situ hybridization from Singer Lab
Added on: 07-Oct-2004 Hits: 1226
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Preparation of sections for in situ hybridization 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 1108
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Preparation of stock solutions for FISH 
Description: Preparation of stock solutions for FISH
Added on: 07-Oct-2004 Hits: 837
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PRETREATMENT OF CHROMOSOME PREPARATIONS  
Description: PRETREATMENT OF CHROMOSOME PREPARATIONS
Added on: 17-Dec-2009 Hits: 309
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QUALITY CONTROL OF CHROMOSOME PREPARATIONS 
Description: QUALITY CONTROL OF CHROMOSOME PREPARATIONS
Added on: 17-Dec-2009 Hits: 326
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REPROBING OF PREPARATIONS 
Description: REPROBING OF PREPARATIONS
Added on: 17-Dec-2009 Hits: 351
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RNA In Situ Hybridization of Dissected Gonads 
Description: RNA In Situ Hybridization of Dissected Gonads with Digoxygenin-Labeled Probes
Added on: 07-Oct-2004 Hits: 782
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Sectioning mouse embryos for mRNA detection 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 763
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Sectioning of whole mount hybridized embryos 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 812 Rating: 1 (1 Vote)
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Single and Double FISH protocols for Drosophila 
Description: This procedure is taken from a recently published book chapter. The reference is: Hughes, S. and Krause, H.M. (1998) Single and double FISH protocols for Drosophila. in: Protocols in confocal microscopy., ed. Stephen Paddock, Humana Press Inc., in press.
Added on: 30-Nov-2004 Hits: 757
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Specific detection of double-stranded DNA sequences in Lowicryl K4M sections by ish 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 726
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SQUASH PREPARATION OF PLANT CHROMOSOMES 
Description: SQUASH PREPARATION OF PLANT CHROMOSOMES
Added on: 17-Dec-2009 Hits: 335
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STRINGENT WASHES 
Description: STRINGENT WASHES
Added on: 17-Dec-2009 Hits: 447
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Subbing of slides for mouse embryo sections for mRNA detection 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 731
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Successive steps for Lowicryl K4M embedding and ultrathin sectioning of somatic mammalian cells prio 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 816
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Successive steps for specific detection of DNA sequences by post-embedding in situ hybridization 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 690
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Successive steps for specific detection of RNA molecules in Lowicryl K4M sections by in situ hybridi 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 708
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Synthesis of digoxigenin or fluorescein labelled RNA probe 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 983
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Synthesis of single-stranded RNA probe for mRNA detection in tissue sections 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 842
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Tavi's Multicolor FISH Page 
Description: Tavi's Multicolor FISH Page (fluorescence in situ hybridization)
Added on: 13-Feb-2009 Hits: 570
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Treatment of slides prior to FISH 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 884
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Two colour in situ hybridization 
Description: Two colour in situ hybridization - sequential alkaline phosphatase staining with chromogenic substrates of zebrafish embryos. Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 1062
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Two colour ISH 
Description: Two colour ISH - sequential alkaline phosphatase staining with chromogenic substrates of chick, mouse, and Xenopus embryos. Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 913
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Whole mount hybridization, washing, and detection of probe (method 1) 
Description: Protocols from Oxford Practical Approach Series.
Added on: 08-Oct-2004 Hits: 1007
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Forums
Last 20 Forum Messages

IHC detection system
Last post by Carl in General Immunohistochemistry on Apr 16, 2012 at 09:51:05

Sections not adhering to slide
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Polyethylene glycol embedded human tissue
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Immuohistochemistry of adipose tissue
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Bubbles forming during hybridization in fresh frozen tissue
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"Far"-IF staining?
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extracellular epitope labeling
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Protocol for IHC B-catenin rat bone?
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Nick Translation
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advice on identifying muscle fiber types on stained image
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iNOS in FFPWS...
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What tissue?
Last post by ole in General Immunohistochemistry on Dec 22, 2011 at 23:20:54

need help wth IHC-P
Last post by Carl in General Immunohistochemistry on Dec 20, 2011 at 20:03:51

Problems with TH staining in frozen human brain
Last post by Carl in General Immunohistochemistry on Dec 09, 2011 at 06:03:52

MOM kit trouble with non specific staining
Last post by Carl in General Immunohistochemistry on Dec 04, 2011 at 16:59:34

Three questions
Last post by salton in General In Situ Hybridization on Dec 02, 2011 at 16:10:42

IHC of spinal cord without PFA perfusion
Last post by Carl in General Immunohistochemistry on Dec 02, 2011 at 08:19:31


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· 1: Anderson Lab In Situ Hybridization Protocols
· 2: Detection of mRNA by in situ hybridization
· 3: DAKO Handbook: Immunohistochemical Staining Methods
· 4: 35S-labeled probe
· 5: DIG-labeled probe
· 6: Detection and amplification of FISH signal
· 7: Chromosome In Situ Hybridization using biotin labeled probes
· 8: In Situ Hybridization
· 9: Chick, mouse, and Xenopus two colour whole mount ISH
· 10: Autoradiography for mRNA detection in mouse embryo tissue sections
 
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