Detection by in-situ fluorescence of short, single-copy sequences of chromosomal DNA

Short single-copy probes have been widely used in plant molecular biology. However, they have rarely been effective in plant research using in situ hybridization techniques, possibly due to limitations imposed by the cell wall. We recently developed two fluorescence in situ hybridization protocols for the single-copy sequence detection in soybean. By enzymatically removing the cell wall, single-copy sequences as short as 1 kb were detected by probes using standard fluorescence in situ hybridization or PCR-primed in situ hybridization (PCR-PRINS). Such technology is useful for genome analysis, in plant molecular, cellular, and biotechnological research.