caa a22 4500 606190597 CHVBK 20210128100856.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s10577-015-9495-3 doi (NATIONALLICENCE)springer-10.1007/s10577-015-9495-3 Identification of the centromeric repeat in the threespine stickleback fish ( Gasterosteus aculeatus ) [Elektronische Daten] [Jennifer Cech, Catherine Peichel] Centromere sequences exist as gaps in many genome assemblies due to their repetitive nature. Here we take an unbiased approach utilizing centromere protein A (CENP-A) chomatin immunoprecipitation followed by high-throughput sequencing to identify the centromeric repeat sequence in the threespine stickleback fish (Gasterosteus aculeatus). A 186-bp, AT-rich repeat was validated as centromeric using both fluorescence in situ hybridization (FISH) and immunofluorescence combined with FISH (IF-FISH) on interphase nuclei and metaphase spreads. This repeat hybridizes strongly to the centromere on all chromosomes, with the exception of weak hybridization to the Y chromosome. Together, our work provides the first validated sequence information for the threespine stickleback centromere. Springer Science+Business Media Dordrecht, 2015 Centromere nationallicence CENP-A nationallicence ChIP-seq nationallicence Gasterosteus aculeatus nationallicence Threespine stickleback nationallicence BAC : Bacterial artificial chromosome nationallicence BSA : Bovine serum albumin nationallicence CEN : Centromeric repeat sequence nationallicence CenH3 : Centromeric histone H3 nationallicence CENP-A : Centromere protein A nationallicence CENP-B : Centromere protein B nationallicence ChIP : Chromatin immunopreciptiation nationallicence ChIP-seq : Chromatin immunopreciptiation sequencing nationallicence DAPI : 4',6'-Diamidino-2-phenlyindole nationallicence dpf : Days post-fertilization nationallicence EDTA : Ethylenediaminetetraacetic acid nationallicence FISH : Fluorescence in situ hybridization nationallicence GacCEN : Threespine stickleback (Gasterosteus aculeatus) centromeric repeat sequence nationallicence H3 : Histone H3 nationallicence HOR : Higher-order repeat nationallicence IF-FISH : Immunofluorescence combined with FISH nationallicence IP : Immunoprecipitation nationallicence KCl : Potassium chloride nationallicence MNase : Micrococcal nuclease nationallicence PBS : Phosphate-buffered saline nationallicence PBST : Phosphate-buffered saline Tween-20 nationallicence PCR : Polymerase chain reaction nationallicence PMSF : Phenylmethanesulfonylfluoride nationallicence POF1 : Pacific Ocean female 1 nationallicence POF2 : Pacific Ocean female 2 nationallicence RPM : Reads per million nationallicence SDS : Sodium dodecyl sulfate nationallicence SDS-PAGE : Sodium dodecyl sulfate polyacrylamide gel electrophoresis nationallicence SSC : Saline-sodium citrate nationallicence 3′ RACE : Rapid amplification of cDNA ends nationallicence Cech Jennifer Divisions of Human Biology and Basic Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave North, Mailstop C2-023, 98109, Seattle, WA, USA aut Peichel Catherine Divisions of Human Biology and Basic Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave North, Mailstop C2-023, 98109, Seattle, WA, USA aut Chromosome Research Springer Netherlands 23/4(2015-12-01), 767-779 0967-3849 23:4<767 2015 23 10577 https://doi.org/10.1007/s10577-015-9495-3 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10577-015-9495-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Cech Jennifer Divisions of Human Biology and Basic Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave North, Mailstop C2-023, 98109, Seattle, WA, USA aut NATIONALLICENCE 700 1- Peichel Catherine Divisions of Human Biology and Basic Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave North, Mailstop C2-023, 98109, Seattle, WA, USA aut NATIONALLICENCE 773 0- Chromosome Research Springer Netherlands 23/4(2015-12-01), 767-779 0967-3849 23:4<767 2015 23 10577