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A very detailed protocol from Dept of Medical Genetics, University Hospital Gent, B-9000 Gent Belgium. ...

50 FISH protocols (Oxford Practical Approach Series) new recommended protocol
There are 50 protocols in this section: Autoradiography for mRNA detection in mouse embryo tissue sections    Protocol Chick, mouse, and Xenopus two colour whole mount in ...

In situ hybridization using oligonucleotide probe new recommended protocol
Protocol for frozen sections with 35S labeled olgonucleotide probe. From GeneDetect.com (PDF version). ...

Fluorescent in situ Hybridization (FISH) protocol
This page maintained by Pasteur Institute briefly presents the introduction to fluorescent in situ hybridization technique (FISH). This technique is used for the detection of target molecules with a system of coupled antibodies and fluorochromes. ...

Single and Double FISH protocols for Drosophila new protocol
From Krause Lab Protocols. This procedure is taken from a book chapter. 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. ...

Fluorescent in situ hybridization (FISH) protocol new protocol
A protocol (manual) from www.cancergenetics.com. Fluorescent in situ hybridization (FISH) protocol. ULSTM-dGreen and ULSTM-Rhodamine labeled probes for translocation, locus specific and chromosome copy number detection. ...

PCR-derived ssDNA Probes for Fluorescent In Situ Hybridization to HIV-1 RNA new protocol
Tech Notes Original Article. Marlyse C. Knuchela et al. Journal of Histochemistry and Cytochemistry, Vol. 48, 285-294, February 2000 Abstract We developed a simple and rapid technique to synthesize single-stranded DNA (ssDNA) probes for fluores ...

In situ Hybridization techniques new protocol
In situ hybridization (ISH) can be used to detect nucleic acid sequences in tissue and cell cultures. Its applications include detection of mRNA expression in situ, chromosomal rearrangement, viral RNA expression in host cell etc. General steps for in situ ...

Anderson Lab In Situ Hybridization Protocols new protocol
These protocols describe non-radioactive methods for in situ hybridization on frozen sections, whole mount embryos and on cultured cells. They have been freely adapted and modified to greater or lesser extents from the protocols of Richard Harland, David Wilki ...

Nonradioactive in situ hybridization protocols new protocol
Cytochemical Detection of mRNA by nonradioactive in situ hybridization A nonradioactive method for in situ hybridization. This page describes the current favorite non-radioactive in situ hybridization t ...

In situ hybridization on frozen sections new protocol
Protocol by Josiah N. Wilcox, Ph.D., Wilcox lab in Winship Cancer Institute of the School of Medicine of Emory University. ...

RNA probe preparation protcol for in situ hybridizaiton. by Josiah N. Wilcox, Ph.D. (based on Melton et al., 1984). Wilcox lab at Emory University. ...

Protocol on tissue section preparation for in situ hybridization. By Dr.Josiah N. Wilcox, Wilcox lab at Emory University. ...

In situ hybridization protocol on embryo tissue sections. Protocol based on Kornberg et al., Cell 40:45, 1985. ...

mRNA In Situ Hybridization with Nick-Translated Probes protocol
Short protocol on In Situ Hybridization. (Spector lab at Cold Spring Harbor Lab). ...

Probe synthesis for in situ hyb new protocol
This is essentially according to the method recommended by Boehringer Mannheim. Cut 20 ug of plasmid DNA with appropriate enzyme in 100 ul. Pheno1, phenol/chloroform, chloroform and ethanol precipitate. Dissolve in 15 ul ddH20.(Hiroyuki Takeda lab, National in ...

Wholemount in situ hybridisations on zebrafish embryos using digoxigenin probes protocol
Protocol by Alex Schier on Wholemount in situ hybridisations on zebrafish embryos using digoxigenin probes. ...

In this example stain with flh in black (NBT substrate) and with gsc in orange (INT substrate). (Kira, National institute of Genetics, Japan) ...

In situ hybridization question troubleshooting
I am doing ISHH using 33P labeled-riboprobes on rat brain tissue and I'm having a problem during the development of the slides. I dip each slide in NBT2 emulsion (diluted 1:1 in water), let dry in the dark standing up for 3 hours and then store in sli ...

More in situ hybridization protocols

fluorescence in situ hybridization (FISH) protocols
(Curr Protoc Hum Genet.)
1. Curr Protoc Hum Genet. 2007 Jan;Chapter 8:Unit 8.8.

Preparation of cells from formalin-fixed, paraffin-embedded tissue for use in
fluorescence in situ hybridization (FISH) experiments.

Weremowicz S, Schofield DE.

Harvard Medical School, Boston, Massachusetts, USA.

Numerical and structural chromosome abnormalities can be accurately detected in
cells from archived tissues using fluorescence in situ hybridization (FISH). This
unit describes two common approaches to performing FISH in formalin-fixed,
paraffin-embedded tissue. The first approach utilizes 4 to 6 microm tissue
sections in cases for which preserving tissue morphology is necessary, and the
second involves extraction of intact nuclei from 50 microm tissue sections. To
interpret FISH results using 4 to 6 microm sections, an adequate number of nuclei
must be evaluated to perform statistical analysis. Evaluation of 30 to 50 nuclei
from the single cell suspension generally gives an interpretable result.

2: Curr Protoc Hum Genet. 2005 May;Chapter 4:Unit 4.3.

In situ hybridization to metaphase chromosomes and interphase nuclei.

Knoll JH, Lichter P.

Children's Mercy Hospital, University of Missouri-Kansas City School of Medicine,
Kansas City, USA.

In situ hybridization is used to determine the chromosomal map location and the
relative order of genes and DNA sequences within a chromosomal band. It can also
be used to detect aneuploidy, gene amplification, and subtle chromosomal
rearrangements. Fluorescence in situ hybridization (FISH), probably the most
widely used method, is described in the first basic protocol. Two support
protocols are provided to amplify weak fluorescent signals obtained in FISH.
Nonisotopic probes can also be detected by enzymatic reactions using horseradish
peroxidase or alkaline phosphatase, as described in alternate protocols.
Nonisotopic labeling of DNA probes by nick translation is described in a support
protocol. The order of closely spaced FISH probes along chromosomes in interphase
nuclei can be determined. A basic protocol for isotopic in situ hybridization
(IISH) with (3)H is provided followed by a support protocol for preparation of
autoradiographic emulsion.

3: Curr Protoc Hum Genet. 2001 May;Chapter 8:Unit 8.9.

Preparation of amniocytes for interphase fluorescence in situ hybridization

Schwartz S, Micale MA, Becker L.

Case Western Reserve University, University Hospitals of Cleveland, Cleveland,
Ohio, USA.

Although FISH has been used to clarify deletions or structural rearrangements,
recent work has focused increasingly on its applications to interphase analysis.
This unit describes preparation of uncultured amniotic fluid cells for FISH
analysis. Cells are swollen, then slides are prepared using either a
cytocentrifuge or standard methods. These are then fixed and permeabilized for
subsequent FISH. An Alternate Protocol describes attachment of amniocytes to a
glass or plastic surface followed by hypotonic swelling, fixation, and
permeabilization for subsequent FISH. Interphase FISH analysis of amniotic fluid
cells is also described.

(Selected in situ hybridization reviews)
1: Brown LA, Huntsman D.
Fluorescent in situ hybridization on tissue microarrays: challenges and
J Mol Histol. 2007 Jan 10; [Epub ahead of print]

2: Jiang J, Gill BS.
Current status and the future of fluorescence in situ hybridization (FISH) in plant genome research.
Genome. 2006 Sep;49(9):1057-68. Review.

3: Liehr T, Starke H, Heller A, Kosyakova N, Mrasek K, Gross M, Karst C,
Steinhaeuser U, Hunstig F, Fickelscher I, Kuechler A, Trifonov V, Romanenko SA, Weise A.
Multicolor fluorescence in situ hybridization (FISH) applied to FISH-banding.
Cytogenet Genome Res. 2006;114(3-4):240-4. Review.

4: Jubb AM, Pham TQ, Frantz GD, Peale FV Jr, Hillan KJ.
Quantitative in situ hybridization of tissue microarrays.
Methods Mol Biol. 2006;326:255-64. Review.

5: Smith MD, Ahern M, Coleman M.
The use of combined immunohistochemical labeling and in situ hybridization to colocalize mRNA and protein in tissue sections.
Methods Mol Biol. 2006;326:235-45. Review.

6: Hewitson TD, Kelynack KJ, Darby IA.
Histochemical localization of cell proliferation using in situ hybridization for histone mRNA.
Methods Mol Biol. 2006;326:219-26. Review.

7: Houben A, Orford SJ, Timmis JN.
In situ hybridization to plant tissues and chromosomes.
Methods Mol Biol. 2006;326:203-18. Review.

8: Owens NC, Hess FM, Badoer E.
In situ hybridization using riboprobes on free-floating brain sections.
Methods Mol Biol. 2006;326:163-71. Review.

9: Hargrave M, Bowles J, Koopman P.
In situ hybridization of whole-mount embryos.
Methods Mol Biol. 2006;326:103-13. Review.

10: Asp J, Abramsson A, Betsholtz C.
Nonradioactive in situ hybridization on frozen sections and whole mounts.
Methods Mol Biol. 2006;326:89-102. Review.

11: Chotteau-Lelievre A, Dolle P, Gofflot F.
Expression analysis of murine genes using in situ hybridization with radioactive and nonradioactively labeled RNA probes.
Methods Mol Biol. 2006;326:61-87. Review.

12: Darby IA, Bisucci T, Desmouliere A, Hewitson TD.
In situ hybridization using cRNA probes: isotopic and nonisotopic detection methods.
Methods Mol Biol. 2006;326:17-31. Review.

13: Tesch GH, Lan HY, Nikolic-Paterson DJ.
Treatment of tissue sections for in situ hybridization.
Methods Mol Biol. 2006;326:1-7. Review.

14: Oliveira AM, French CA.
Applications of fluorescence in situ hybridization in cytopathology: a review.
Acta Cytol. 2005 Nov-Dec;49(6):587-94. Review.

15: Hicks DG, Longoria G, Pettay J, Grogan T, Tarr S, Tubbs R.
In situ hybridization in the pathology laboratory: general principles, automation, and emerging research applications for tissue-based studies of gene
J Mol Histol. 2004 Aug;35(6):595-601. Review.

16: Henke RT, Maitra A, Paik S, Wellstein A.
Gene expression analysis in sections and tissue microarrays of archival tissues by mRNA in situ hybridization.
Histol Histopathol. 2005 Jan;20(1):225-37. Review.

17: Ransick A.
Detection of mRNA by in situ hybridization and RT-PCR.
Methods Cell Biol. 2004;74:601-20. Review.

18: Bartlett JM.
Fluorescence in situ hybridization: technical overview.
Methods Mol Med. 2004;97:77-87. Review.

19: Schwarzacher T.
DNA, chromosomes, and in situ hybridization.
Genome. 2003 Dec;46(6):953-62. Review.

20: Levsky JM, Singer RH.
Fluorescence in situ hybridization: past, present and future.
J Cell Sci. 2003 Jul 15;116(Pt 14):2833-8. Review.

21: Qian X, Lloyd RV.
Recent developments in signal amplification methods for in situ hybridization.
Diagn Mol Pathol. 2003 Mar;12(1):1-13. Review.

22: Muller F.
Processing retinal tissue for in situ hybridization.
Int Rev Neurobiol. 2002;47:85-92. Review.

23: Laurie DJ, Schrotz PC, Monyer H, Amtmann U.
Processing rodent embryonic and early postnatal tissue for in situ hybridization with radiolabelled oligonucleotides.
Int Rev Neurobiol. 2002;47:71-83. Review.

24: Wisden W, Morris BJ.
In situ hybridization with oligonucleotide probes.
Int Rev Neurobiol. 2002;47:3-59. Review.

25: Ariza-McNaughton L, Krumlauf R.
Non-radioactive in situ hybridization: simplified procedures for use in whole-mounts of mouse and chick embryos.
Int Rev Neurobiol. 2002;47:239-50. Review.

26: Augood SJ, McGowan EM, Finsen BR, Heppelmann B, Emson PC.
Non-radioactive in situ hybridization using alkaline phosphatase-labelled oligonucleotides.
Int Rev Neurobiol. 2002;47:173-201. Review.

27: Gundlach AL, O'Shea RD.
Quantitative analysis of in situ hybridization histochemistry.
Int Rev Neurobiol. 2002;47:135-70. Review.

28: Nicholson LF.
Processing human brain tissue for in situ hybridization with radiolabelled oligonucleotides.
Int Rev Neurobiol. 2002;47:105-16. Review.

In situ hybridization technical books
(Biowww Bookshelf)
In situ hybridization methods

elute proteins fom PAGE gel
elude 24KD protein from native PAGE gel without loss of activity

Zebrafish in situ hybridization
with AP-conjugated Antibody and NBT/BCIP

RNA in situ hybridization
with DIG Labelled probes, AP-conjugated Ab and NBT/BCIP

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