Efficient isolation method for high-quality genomic DNA from cicada exuviae

Ecology and Evolution
Hoa Quynh NguyenYikweon Jang

Abstract

In recent years, animal ethics issues have led researchers to explore nondestructive methods to access materials for genetic studies. Cicada exuviae are among those materials because they are cast skins that individuals left after molt and are easily collected. In this study, we aim to identify the most efficient extraction method to obtain high quantity and quality of DNA from cicada exuviae. We compared relative DNA yield and purity of six extraction protocols, including both manual protocols and available commercial kits, extracting from four different exoskeleton parts. Furthermore, amplification and sequencing of genomic DNA were evaluated in terms of availability of sequencing sequence at the expected genomic size. Both the choice of protocol and exuvia part significantly affected DNA yield and purity. Only samples that were extracted using the PowerSoil DNA Isolation kit generated gel bands of expected size as well as successful sequencing results. The failed attempts to extract DNA using other protocols could be partially explained by a low DNA yield from cicada exuviae and partly by contamination with humic acids that exist in the soil where cicada nymphs reside before emergence, as shown by spectroscopic measurements....Continue Reading

References

Jan 18, 2003·Journal of Microbiological Methods·Michael D BraidChristopher L Kitts
Mar 27, 2007·Journal of Insect Physiology·Songkun SuJürgen Tautz
Feb 8, 2011·Nucleic Acids Research·Claudia BaarPhilipp Holliger
Jul 4, 2012·Journal of Applied Microbiology·C SchraderR Johne

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BETA
PCR
electrophoresis

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SPSS

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