IN SITU HYBRIDIZATION PROTOCOLS
The first three protocols describe isolation and purification of probe (plasmid) and plant genomic DNA, and the preparation of genomic blocking DNA. Other protocols describe details of chromosome preparation and in situ hybridization . The last protocol describes a modified N-banding technique that can be combined with in situ hybridization for simultaneous chromosome identification and mapping.
Biotin labeling and hybridization of probe to chromosomes
Chromosome preparation for in situ hybridization
Detection of ISH signals with fluorochromes
Detection of ISH signals with streptavidin
Isolation of plasmid (probe) DNA for in situ hybridization
Isolation of plant genomic DNA for GISH
Modified N-banding technique for sequential chromosome banding and ISH analysis
Preparation of genomic blocking DNA
Purification of the biotin-labeled probe
