Comparison of the Effectiveness of Different Dna Isolation Methods and Methodological Modifications in Molted Feathers of Pigeons and Nightingales


Şentürk N., Çobanoğlu Ö., Taşkın S. D., Ardıçlı S.

12th International Medicine and Heath Sciences Researches Congress, Ankara, Turkey, 18 - 19 March 2023, vol.12, no.78, pp.515-516

  • Publication Type: Conference Paper / Summary Text
  • Volume: 12
  • City: Ankara
  • Country: Turkey
  • Page Numbers: pp.515-516
  • Bursa Uludag University Affiliated: Yes

Abstract

Non-invasive sampling methods are major constituents for the genetic analysis of birds. Thus, molted feathers are a precious source of DNA. For successful genetic assessments, obtaining high-quantity and -quality DNA samples plays a vital role. Therefore, this study aimed to compare different DNA extraction methods to achieve higher amounts and purity levels from molted feathers of pigeons (Columba livia f. domestica) and nightingales (Luscinia megarhynchos). We evaluated 55 animals consisting of 52 pigeons and three nightingales. We performed three commercially available DNA extraction kits and a rapid alkaline extraction method. We next compared the combination of simple alterations comprised of increased or decreased incubation time, different parts of the feathers, and modification of temperature and volume of lysis buffer to increase DNA yield and quality. DNA concentration (ng/mL) and absorbance ratio at 260/280 were measured using a Nanodrop spectrophotometer. Kruskal-Wallis or the Mann-Whitney U tests were performed for the statistical comparisons. The duration of lysis with the proteinase K incubation, including overnight or 3h, did not affect the yield and quality. The anatomical regions for the cross-sections through the feather significantly impacted the DNA absorbance ratio (P<0.05). In this context, the feather samples from only the calamus, including the superior and inferior umbilicus, exhibited better results than those with barbs. The samples with only the calamus had higher DNA yield than those with the inferior rachis (without vanes) (P<0.05). The DNA quality was much better in commercial kits compared to the alkaline extraction method. DNA kit brand influenced the yield (P<0.05) but not quality (P>0.05). Anatomical differences in birds' feathers from different species cause differences in sample preparation before the lysis step. The modifications reported in this study may be helpful for the DNA extraction from the feather samples collected non-invasively in the field for genetic analysis in birds.