CHROMOSOME MICRODISSECTION AND REGION-SPECIFIC LIBRARIES FROM PACHYTENE CHROMOSOMES OF MAIZE (ZEA-MAYS L.)

For the first time, pachytene chromosomes have been used for micro-dis section, amplification and cloning using the short arm of maize chromo some 6. For this purpose, methods for the preparation of unstained pac hytene chromosomes were developed. In two separate experiments, 10 and 12 segments of the satellite region were isolated utilizing glass nee dles and DNA was amplified by linker-adaptor-PCR. After amplification, maize-specific DNA was verified by genomic Southern hybridizations. C hromosomal in situ suppression hybridization confirmed that the adapto r PCR products originated from the micro-dissected 6S region. The adap tor PCR products were cloned into a plasmid vector and the chromosome region-specific libraries were characterized.