Efficiency and Reliability of DNA Extraction Methods for Ixodid Ticks (Acari: Ixodidae): A Comparative Tour

Abstract

ABSTRACT
It is well understood that DNA extraction is the primary as well as the crucial step while studying ticks at molecular level. The DNA extraction methods should be straightforward, and enough efficient as well as reliable in terms of DNA quality and quantity. In the present study, two non-commercial methods named phenol chloroform isoamyl alcohol (PCI) and cetyl-trimethyl ammonium bromide (CTAB) and one commercial DNeasy tissue kit were compared based on enzymatic digestion followed by the steps of DNA extraction. Nanodrop spectrophotometer was used to assess the quality of DNA. Further, the absence of PCR inhibitors and the DNA quality were evaluated by PCR amplification of the tick mitochondrial cytochrome C oxidase subunit I (COI) gene using tick-specific primers. The absorbance ratio for all the methods ranges from 1.7–2.05. DNA pellets were found light to dark brown in PCI method, while it was clear in CTAB and kit method. A fragment of ⁓500bp was successfully amplified with all three methods, except one sample which was extracted with kit method. Thus, the conclusion is extended in the form of suitability, efficiency as well as reliability of P CI method followed by CTAB method in extraction of DNA from ticks’ samples.

Keywords: ticks, DNA, extraction, absorbance, metagenomics, PCR

Cite this Article: Tripti Negi,∙Kusum Arunachalam. Efficiency and Reliability of DNA Extraction Methods for Ixodid Ticks (Acari: Ixodidae): A Comparative Tour. International Journal of Animal Biotechnology and Applications. 2019; 5(1 & 2): 1–9p.