By Joel Rothman, Andrew Singson
The moment a part of an up-to-date version of the vintage Methods in phone Biology, Volume forty eight, this e-book emphasizes diversified tools and applied sciences had to examine C. elegans, either as an built-in organism and as a version procedure for study inquiries in phone, developmental, and molecular biology, in addition to in genetics and pharmacology. through directing its viewers to tried-and-true and state of the art recipes for learn, this accomplished assortment is meant to lead investigators of C. elegans for years to come.
- Diverse, updated concepts lined could be important to the broadening neighborhood of C. elegans researchers for years to come
- Chapters written through leaders within the field
- Tried and actual tools bring busy researchers a one-stop compendium of crucial protocols
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Extra resources for Caenorhabditis elegans : cell biology and physiology
Maddox and Paul S. _4)TD$IG] Fig. 4 The steps and tools for worm dissection for mounting and live imaging of early embryos. (A) Gravid adult held by #3 forceps (left), to be cut by hypodermic needle (right). (B) Freed embryos and worm carcass being aspirated by pulled glass mouth pipette (bottom right). (C) Eyelash tool being used to sweep embryos that have been transferred to an agarose pad. (D) Notch has been cut from the agarose pad to indicate the location and orientation of the collection of embryos aligned with the eyelash tool.
2007). , 2008). In summary, immunofluorescence-based techniques provide a powerful repertoire of tools for tackling biological problems in C. elegans. Immunolocalization also complements other molecular studies of gene expression. Whereas Northern blots and RNA in situ hybridization studies (Seydoux and 2. Immunofluorescence Microscopy 41 Fire, 1995) provide important information on the transcriptional regulation of particular genes, protein immunofluorescence reveals where the majority of the endogenous protein is actually localized and presumably functions.
In some instances, however, it is necessary to preadsorb the secondary fluorescent antibody with fixed nematodes (see Section III, below) to reduce background staining. Antibodies are notoriously sensitive to repeated freeze-thaw cycles. As a result, antibody stocks should be aliquoted prior to storage at À 20 or À 80 C. Alternatively, many antibodies stocks can be mixed 1:1 with 100% glycerol as a way of keeping a nonfrozen À20 C stock. 04% sodium azide to prevent bacterial growth. 44 Diane C.