Chromosome micro-dissection and region-specific libraries from pachytene chromosomes of maize (Zea mays. L.)

Abstract

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