TY - JOUR
T1 - Quantitative localization of chromosomal loci by immunofluorescence
AU - Brickner, Donna Garvey
AU - Light, William
AU - Brickner, Jason H.
N1 - Funding Information:
The authors thank John Sedat for sharing the GFP-Lac repressor and the LacO array plasmids, Francoise Stutz for the GAL2 LacO plasmid, Kevin Redding for developing the immunofluorescence protocol on which these methods are based and members of the Brickner laboratory for helpful comments on this chapter. J. H. B. is supported by NIH grant GM080484, the W. M. Keck Foundation and the Baldwin Fund for Biomedical Research.
PY - 2010
Y1 - 2010
N2 - DNA within the yeast nucleus is spatially organized. Yeast telomeres cluster together at the nuclear periphery, centromeres cluster together near the spindle pole body, and both the rDNA repeats and tRNA genes cluster within the nucleolus. Furthermore, the localization of individual genes to subnuclear compartments can change with changes in transcriptional status. As such, yeast researchers interested in understanding nuclear events may need to determine the subnuclear localization of parts of the genome. This chapter describes a straightforward quantitative approach using immunofluorescence and confocal microscopy to localize chromosomal loci with respect to well characterized nuclear landmarks.
AB - DNA within the yeast nucleus is spatially organized. Yeast telomeres cluster together at the nuclear periphery, centromeres cluster together near the spindle pole body, and both the rDNA repeats and tRNA genes cluster within the nucleolus. Furthermore, the localization of individual genes to subnuclear compartments can change with changes in transcriptional status. As such, yeast researchers interested in understanding nuclear events may need to determine the subnuclear localization of parts of the genome. This chapter describes a straightforward quantitative approach using immunofluorescence and confocal microscopy to localize chromosomal loci with respect to well characterized nuclear landmarks.
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U2 - 10.1016/S0076-6879(10)70022-7
DO - 10.1016/S0076-6879(10)70022-7
M3 - Article
C2 - 20946825
AN - SCOPUS:77956039093
SN - 0076-6879
VL - 470
SP - 569
EP - 580
JO - Methods in Enzymology
JF - Methods in Enzymology
IS - C
ER -