Immunomagnetic isolated cells were spotted onto poly-L-lysine-coated coverslips and left to settle for 15–30 minutes at RT. Cells were then processed for immune-DNA FISH as described previously10 (link), with the following modifications. Digoxigenin (DIG)-labeled probes were prepared by labeling 1 μg of BAC DNA using DIG-Nick Translation Mix (Sigma-Aldrich) and cells were subjected to IF with an anti-DIG antibody (Sigma-Aldrich). The following BAC probes from BACPAC resources, CHORI were used: Cav2 BAC (RP23-448I4), cJun BAC (RP24-282B18), and Nup62 BAC (RP23-403D23).
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Immuno-DNA FISH Analysis of Genomic Loci
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Other organizations : Discovery Institute, Sanford Burnham Prebys Medical Discovery Institute
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Variable analysis
independent variables
- Immunomagnetic isolated cells
- Digoxigenin (DIG)-labeled probes prepared by labeling 1 μg of BAC DNA using DIG-Nick Translation Mix (Sigma-Aldrich)
- BAC probes from BACPAC resources, CHORI: Cav2 BAC (RP23-448I4), cJun BAC (RP24-282B18), and Nup62 BAC (RP23-403D23)
- Immune-DNA FISH as described previously
- Poly-L-lysine-coated coverslips
- Cells were left to settle for 15–30 minutes at RT
- Cells were subjected to IF with an anti-DIG antibody (Sigma-Aldrich)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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