Chromosome numbers and karyotypes were analysed as described by Jang et al. [15 (link),20 (link)] using standard Feulgen staining. Chromosomal spreads for FISH and GISH were prepared by enzymatic digestion and squashing, as described in Jang et al. [15 (link),20 (link)].
Probes used for FISH were: satellite DNA PaB6 isolated from the B6 genome in plasmid pGEM-T easy [28 (link)], 35S rDNA (18S/25S rDNA) from Arabidopsis thaliana in plasmid pSK+, and 5S rDNA from Melampodium montanum (Asteraceae) in plasmid pGEM-T easy, labeled with biotin or digoxygenin (Roche, Vienna, Austria). Probes were labeled either directly by PCR (5S rDNA and satellite DNA PaB6) or using a nick translation kit (35S rDNA; Roche). A commercially available, directly Cy3-labelled PNA (peptide nucleic acid) probe to vertebrate telomeric sequences (CCCTAA)3 (Dako, Glostrup, Denmark) was used as described in [10 (link)]. FISH was performed as described in Jang et al. [15 (link),20 (link)].
Probes used for FISH were: satellite DNA PaB6 isolated from the B6 genome in plasmid pGEM-T easy [28 (link)], 35S rDNA (18S/25S rDNA) from Arabidopsis thaliana in plasmid pSK+, and 5S rDNA from Melampodium montanum (Asteraceae) in plasmid pGEM-T easy, labeled with biotin or digoxygenin (Roche, Vienna, Austria). Probes were labeled either directly by PCR (5S rDNA and satellite DNA PaB6) or using a nick translation kit (35S rDNA; Roche). A commercially available, directly Cy3-labelled PNA (peptide nucleic acid) probe to vertebrate telomeric sequences (CCCTAA)3 (Dako, Glostrup, Denmark) was used as described in [10 (link)]. FISH was performed as described in Jang et al. [15 (link),20 (link)].
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