DeSalle et al. 2005

DeSalle R, Gregory TR, Johnston JS. 2005. Preparation of samples for comparative studies of arthropod chromosomes: visualization, In Situ hybridization, and genome size estimation. Methods in Enzymology 395: 460-488.

This paper appears to be formatted like a book chapter, and covers a broad range of topics in arthropod cytogenetics. These authors describe the fundamental concepts of arthropod chromosome research and genome size, focusing primarily on general techniques and best-practices recommendations.

The section concerning chromosome manipulation includes general instructions for preparing chromosome squashes and descriptions of some of the most widely-used stains. Most of the stains described are useful for studying GC content, though silver stains are mentioned for studies of Nucleolus Organizer Regions (NORs), and Flourescent In-Situ Hybridization (FISH) is also described. A table lists books and websites for more detailed protocols.

The section on genome size estimation is obviously of greater direct relevance for my work. Besides strong justification for the work I do, this part of this paper provides some interesting factoids about the range of genome sizes in various arthropod groups, though I know some more recent additions to the database extend these ranges. Genome size estimation protocols, for both Feulgen Image Analysis Densitometry and Flow Cytometry are also provided, including a detailed FC protocol in Box III. Interestingly, the FC protocol includes recommendations for both very long stain incubations (up to 24 hours, but always at least 1h) and very long sampling runs (up to 20 minutes), the latter due to the generally low cell density of insect preparations. I have copied Box III to a MS-Word document for inclusion in my lab notebook.