Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/80848
Title: Immunofluorescence protects RNA signals in simultaneous RNA–DNA FISH
Authors: Lai, Lan-Tian
Lee, Pin Jie
Zhang, Li-Feng
Keywords: Simultaneous RNA–DNA FISH
X chromosome inactivation (XCI)
Terra
Fluorescence in situ hybridization (FISH)
Xist
Issue Date: 2013
Source: Lai, L.-T., Lee, P. J., & Zhang, L.-F. (2013). Immunofluorescence protects RNA signals in simultaneous RNA–DNA FISH. Experimental Cell Research, 319(3), 46-55.
Series/Report no.: Experimental Cell Research
Abstract: Cell research often requires combinational detection of RNA and DNA by fluorescence in situ hybridization (RNA–DNA FISH). However, it is difficult to preserve the fragile RNA signals through the harsh conditions used to denature the DNA template in DNA FISH. The current protocols of RNA –DNA FISH still cannot work robustly in all experiments. RNA–DNA FISH remains as a technically challenging and tedious experiment. By incorporating protein components into the signal detection steps of RNA FISH, which is then followed by a post-fixation step, we established an improved protocol of RNA–DNA FISH. The established method worked satisfyingly and robustly in our studies on Xist (inactivated X chromosome specific transcript) RNA and Terra (telomeric repeat-containing RNA). Our results provided the direct evidence to show that, not all the telomeres are associated with Terra, and a significant fraction of Terra foci do not overlap with telomere DNA in interphase cell nuclei. The improved method of simultaneous RNA–DNA FISH is reliable and time-efficient. It can be used in a variety of biological studies.
URI: https://hdl.handle.net/10356/80848
http://hdl.handle.net/10220/38893
ISSN: 0014-4827
DOI: 10.1016/j.yexcr.2012.11.009
Rights: © 2012 Elsevier Inc. All rights reserved. This is the author created version of a work that has been peer reviewed and accepted for publication by Experimental Cell Research, Elsevier Inc. It incorporates referee’s comments but changes resulting from the publishing process, such as copyediting, structural formatting, may not be reflected in this document. The published version is available at: [http://dx.doi.org/10.1016/j.yexcr.2012.11.009]
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SBS Journal Articles

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