Metaphase chromosome spreads obtained from WT and mutant BRDW1 oocytes were subjected to DNA-FISH analysis using an FITC-conjugated telomeric probe (Biosynthesis, Inc.), as described previously (Baumann et al., 2010 (link)). In brief, surface spread chromosomes were denatured in 70% formamide (VWR International) in 2× SSC at 85°C for 10 min and subsequently chilled in ice-cold 70% ethanol for 5 min. The telomere probe was denatured for 10 min at 85°C and incubated at 37°C for 1 h. Overnight hybridization was performed in a humidified chamber at room temperature, and stringency washes were conducted in a solution containing 50% formamide in 2× SSC as described previously (De La Fuente et al., 2004 (link)).
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Telomere Visualization in Oocyte Chromosomes
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Corresponding Organization :
Other organizations : Cornell University, New York State College of Veterinary Medicine, University of Georgia, Jackson Laboratory
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Variable analysis
independent variables
- Genotype (WT vs. mutant BRDW1)
- Telomere characteristics (e.g., telomere length, structure) in metaphase chromosome spreads
- Experimental procedures (e.g., chromosome spreading, DNA-FISH, hybridization conditions)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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