Fluorescence in situ hybridization was performed as described [18 (link)]. The fic-1 smFISH probes (Biosearch Technologies, Inc) were conjugated to Cy5 fluorophores using the Amersham Cy5 Mono-reactive Dye pack (GE Healthcare). Probe sequences are available upon request. Images in Fig 4 are maximum intensity projections of Z-stacks processed with the FFT Bandpass Filter operations in the image processing program Fiji [47 (link)].
Amersham cy5 mono reactive dye pack
Manufactured by GE Healthcare
The Amersham Cy5 Mono-reactive Dye pack is a fluorescent labeling reagent used in various life science applications. It is designed to covalently attach to biomolecules such as proteins, nucleic acids, and other macromolecules. The dye pack emits fluorescence in the far-red/near-infrared region of the spectrum, which can be detected using appropriate instrumentation.
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2 protocols using amersham cy5 mono reactive dye pack
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Fluorescence in situ Hybridization of fic-1
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Fluorescence In Situ Hybridization Assay
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Fluorescence in situ hybridization was performed as described [58] (link). The gcn-1 and abcf-3 probes (Biosearch Technologies, Inc) were conjugated to the fluorophore Cy5 using the Amersham Cy5 Mono-reactive Dye pack (GE Healthcare). DNA was visualized using 4′,6-diamidino-2-phenylindole (DAPI). The probe sequences used are shown in Tables S3 and S4 . Figures 4D and H are maximum intensity projections of a Z-stack of images processed with the FFT Bandpass Filter operations in the image processing program Fiji. Oligonucleotides used for gcn-1 and abcf-3 FISH probe were described in Table S6 and S7 ).
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