Abstract
Preparing nuclei is necessary in a variety of experimental paradigms to study nuclear processes. In this protocol, we describe a method for rapid preparation of large number of relatively pure nuclei from Ascaris embryos or tissues that are ready to be used for further experiments such as chromatin isolation and ChIP-seq, nuclear RNA analyses, or preparation of nuclear extracts (Kang et al., 2016; Wang et al., 2016).
Keywords: Nuclei isolation, Embryos, Tissue, Nematodes, Ascaris
Background
Nuclei isolation is often the first step in studying the molecular and biochemical aspects of nuclear events. Several methods have been developed to isolate nuclei from different tissues and cell types. However, few nuclei isolation protocols from nematodes other than C. elegans have been described (Ooi et al., 2010; Zanin et al., 2011; Haenni et al., 2012). Embryos of the parasitic nematode Ascaris have been used to prepare a variety of extracts for in vitro cell-free systems (Cohen et al., 2004; Lall et al., 2004). However, these extracts were typically whole cell extracts. Here we describe a method for preparation of nuclei from the nematode Ascaris.
Materials and Reagents
Equipment
Procedure
Notes
Recipes
Acknowledgments
This work was supported in part by NIH grants to R.E.D. (AI0149558 and AI114054).
References
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