Chromosome Spread Immunofluorescence Analysis

Chromosome spreading was performed as described previously [87 (link)]. The slides carrying the spreads were treated with appropriate primary and secondary antibody combinations prior to fluorescence microscopy. The primary antibodies, mouse anti-Myc (1:300), rat anti-HA (3F10; 1:300), and mouse anti-GFP (1:300), were purchased from Roche (Germany). AlexaFluor488-conjugated goat anti-mouse antibody (1:200) and TRITC-conjugated goat anti-rat antibody (1:200) were obtained from Jackson ImmunoResearch Laboratories.

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Ma C.H., Kumar D., Jayaram M., Ghosh S.K, & Iyer V.R. (2023). The selfish yeast plasmid exploits a SWI/SNF-type chromatin remodeling complex for hitchhiking on chromosomes and ensuring high-fidelity propagation. PLOS Genetics, 19(10), e1010986.

Publication 2023

mouse anti-Myc rat anti-HA (3F10; mouse anti-GFP AlexaFluor488-conjugated goat anti-mouse antibody

Anti antibody Antibodies Chromosome Fluorescence antibody microscopy Goat Mouse Tritc

Corresponding Organization : Indian Institute of Technology Bombay

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Variable analysis

independent variables

Primary antibodies: mouse anti-Myc, rat anti-HA, mouse anti-GFP

dependent variables

Fluorescence microscopy of chromosome spreads

control variables

Chromosome spreading procedure as described previously [87]
Antibody concentrations: mouse anti-Myc (1:300), rat anti-HA (3F10; 1:300), mouse anti-GFP (1:300)
Secondary antibodies: AlexaFluor488-conjugated goat anti-mouse (1:200), TRITC-conjugated goat anti-rat (1:200)

controls

Positive control: not mentioned
Negative control: not mentioned

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