Practical in situ Hybridization is aimed at researchers who wish to use efficient and reliable protocols in their work and those who need to check the validity and interpretation of published data. This book presents a reliable, tried-and-tested approach to the methodologies in current use in many laboratories. This online edition is presented in "Standard format" and "Printer-friendly format".

• DNA: DNA in situ hybridization

• PCR Labeling of cloned DNA sequences

• Ethanol Precipitation

• Checking Biotin and Digoxigenin labeled Probe

• Preparation of Chromosome Spreads

• Preparation of Acid-cleaned slides

• Accumulation & fixation of Plant metaphase chromosomes

• Squash preparation of plant chromosomes

• In situ Hybridization data sheet

• Mitotic chromosome preparations from mammalian suspension cell cultures

• Quality control of chromosome preparations

• Pretreatment of chromosome preparations

• Hybridization mixture for cloned and genomic probes

• Denaturation and Hybridization

• Reprobing of preparations

• Strigent washes

• Fluorescent detection of hybridization sites

• Fluorescent staining of DNA and mounting

• Appendix - Buffers and other solutions

Molecular Methods (Laboratory Protocols)

• Restriction digestions and agarose gel electrophoresis

• BAC clones - growth and small-scale isolation methods

• Southern hybridization

• DNA Isolation/Extraction

• DNA/RNA together isolation method

• Agar plates for bacterial plasmid clone selection with ampicilling and GUS

• Acrylamide gel electrophoresis for SSR/microsatellite methods

• Handling, dilution and storage of oligonucleotides

• PCR labelling / labeling

Additional protocols in molecular cytogenetics

This section of the website gives additional protocols, frequently asked questions not included in the book, and additional information or updates.

• Flow Cytometry

• DNA Combing / DNA Fiber preparation  

• Silver Staining of NORs 

• Probe Labeling  Chromosome Preparation Plant root/bud fixation 

• Antifade Mounting Solution 

• Immunostaining methylcytosine on chromosomes  

• Immunostaining of chromosomal/cellular proteins 

• APES-SILANE coating of slides

• Chromic Acid Treatment of Slides

• Alkali hydrolysis of BAC DNA before random-primer labeling

• Frequently Asked Questions

• In situ hybridization data form (Word version)

Spreadsheets (downloads in Excel format): Calculation of Stringency
Calculation of dinucleotide frequencies and comparison with random expectation spreadsheet