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