BioProtocols--nonradioactive in situ hybridization application manual

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Nonradioactive In Situ Hybridization Application Manual (Roche)

http://www.rocheappliescience.com/PROD_INF/MANUALS/InSitu/InSi_toc.htm

Chapter 1 General Introduction to In Situ Hybridization	General Introduction to In Situ Hybridization	9
	Introduction to Hapten Labeling and Detection of Nucleic Acids	11
	Choosing the Right Labeling Method for your Hybridization Experiment	15

Chapter 2 Guidelines for In Situ Hybridization	Guidelines for In Situ Hybridization	21
Chapter 2 Guidelines for In Situ Hybridization	Details of the Technique	22

Chapter 3
Nucleic Acid Hybridization - General Aspects

Nucleic Acid Hybridization - General Aspects

33

Chapter 4 Procedures for Labeling DNA, RNA, and Oligonucleotides with DIG, Biotin, or Fluorochromes	Procedures for Labeling DNA, RNA, and Oligonucleotides with DIG, Biotin, or Fluorochromes	41
	Random primed labeling of ds DNA with DIG-, Biotin- or Fluorescein-High Prime reaction mix	41
	PCR labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes	43
	Nick-translation labeling of ds DNA with Nick Translation Mixes for in situ Probes	49
	Nick-translation labeling of ds DNA with DIG-, Biotin-, or Fluorochrome-labeled dUTP	52
	RNA labeling by in vitro transcription of DNA with DIG, Biotin or Fluorescein RNA Labeling Mix	54
	Oligonucleotide 3'-end labeling with DIG-ddUTP, Biotin-ddUTP, or Fluorescein-ddUTP	58
	Oligonucleotide tailing with a DIG-dUTP, Biotin-dUTP, or Fluorescein-dUTP mixture	60
	Chem-Link labeling of DNA or RNA with DIG or Biotin	63
	Estimating the Yield of DIG-labeled Nucleic Acids	65
	Purification of labeled probes using the High Pure PCR Product Purification Kit	71

Chapter 5 Procedures for In Situ Hybridization to Chromosomes, Cells, and Tissue Sections	Procedures for In Situ Hybridization to whole chromosomes	77
	In situ hybridization to human metaphase chromosomes using DIG-, biotin-, or fluorochrome-labeled DNA probes and detection with fluorochrome conjugates	78
	Detection of chromosomal imbalances using DOP-PCR and comparative genomic hybridization (CGH)	95
	Fluorescence in situ hybridization of a repetitive DNA probe to human chromosomes in suspension	104
	A simplified and efficient protocol for nonradioactive in situ hybridization to polytene chromosomes with a DIG-labeled DNA probe	108
	Multiple-target DNA in situ hybridization with enzyme-based cytochemical detection systems	112
	DNA in situ hybridization with an alkaline phosphatase-based fluorescent detection system	124
	Combined DNA in in situ hybridization and immunocytochemistry for the simultaneous detection of nucleic acid sequences, proteins, and incorporated BrdU in cell preparations	127
	In situ hybridization to mRNA in in vitro cultured cells with DNA probes	135
	Identification of single bacterial cells using DIG-labeled oligonucleotides	139
	Detection of HPV11 DNA in paraffin-embedded laryngeal tissue with a DIG-labeled DNA probe	143
	Detection of mRNA in tissue sections using DIG-labeled RNA and oligonucleotide probes	149
	Detection of mRNA on paraffin-embedded material of the central nervous system with DIG-labeled RNA probes	164
	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	172
	Detection of neuropeptide mRNAs in tissue sections using oligonucleotides tailed with fluorescein-12-dUTP or DIG-dUTP	178
	RNA in situ hybridization using DIG-labeled cRNA probes	181
	Detection of mRNAs on cryosections of the cardiovascular system using DIG-labeled RNA probes	189
	Molecular and Biochemical Analysis of Arabidopsis	197
	Whole mount in situ hybridization for the detection of mRNA in Drosophila embryos	208
	Detection of even-skipped transcripts in Drosophila embryos with PCR/DIG-labeled DNA probe	216
	Whole mount fluorescence in situ hybridization (FISH) of repetitive DNA sequences on interphase nuclei of the small cruciferous plant Arabidopsis thaliana	220

Chapter 6

Ordering Information

231

Chapter 7

Index

237

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