Molecular Methods

There are some practical considerations one should keep in mind when isolating DNA, RNA and protein from ants. Queens tend to be very high in fat content before their mating flight as well as in their physogastric form. Supplementary extractions with ether to remove this fat can be helpful for DNA and RNA isolation. Ants also tend to have much harder exoskeleton and higher surface-to-mass ratio than Drosophila, and complete homogenization of tissue can be difficult without a bead-type shaker homogenizer.

Some experiments are better conducted on dissected tissues or body sections. Physogastric queens lay large quantities of eggs and the mRNA profiles of the queens can become dominated by that of the eggs. Males contain a large amount of sperm and subsequently large amounts of DNA for their body weight. Beginning a DNA extraction with dissected vas deferens containing sperm represents an immediate 10-fold purification. The formic acid in Formicine ants can also be problematic as was found in protein preparations from L. niger workers (Parker et al., 2004a). The acidity can effect isolation and gel loading buffers as well as interfere with enzyme activity assays. The efficiency of phenol extractions in DNA isolations can also be adversely affected by the acid.

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