Journal of Biotechnology Research ISSN: 2413-3256 Vol. 1, No. 4, pp: 16-20, 2015 URL: http://arpgweb.com/?ic=journal&journal=16&info=aims *Corresponding Author 16 Academic Research Publishing Group Comparison of Three Methods of DNA Extraction from Parachipteria willmanni (Acari: Oribatida) Collected in Turkey Sedat Per * Department of Biology, Faculty of Arts and Science, Bozok University, 66900, Yozgat-Turkey Fahriye Sümer Ercan Department of Genetic and Bioengineering, Faculty of Architecture and Engineering, Ahi Evran University, 40200, Kırşehir-Turkey 1. Introduction The ordo Oribatida (Acari) members are saprophagous microarthropods that generally lived in soil ecosystems worldwide [1]. They are the important part of the soil ecosystems and readily sampled in large numbers [2]. Characterization of Acari species is primarily based on morphological differences. Nevertheless, molecular analysis within and between mite species would better clarify their phylogenetic diversity because of their small size [3]. These studies require high quality DNA and so, choosing the most effective DNA isolation protocol is the foremost step before starting the study. Because the prime problem of DNA extraction from small specimen is its quality [4]. Besides, morphologically cryptic species often occur in mite groups [5]. Extraction and purification of DNA is an essential step before starting a study belongs to a molecular biology. In molecular studies, isolated DNA from target sample must be pure enough for PCR amplification and also yield of DNA must be sufficient. Several soil or sediment samples like Acari, contain extremely high amounts of polysaccharides, proteins, and tannins that would interfere with the DNA isolation protocols [6]. The main problem in the isolation and purification of DNA from samples contains degradation of DNA by endonucleases. Recent studies have proposed DNA isolation methods of different mite species [3, 6-9]. Although, there are many studies on identification of this group by classical methods [10-15], molecular-based studies have not found from Turkey. The aim of this research contains improvement a method to isolate DNA from Parachipteria willmanni. Therefore, we compared three different DNA isolation procedures and we made RAPD-PCR with extracted DNA from these methods. This is the first report on the comparison of DNA isolation methods from P. willmanni in Turkey. 2. Materials and Methods 2.1. Sample Collection and Identification Parachipteria willmanni belongs to the family Achipteriidae (Acari: Oribatida), was collected in moss and lichen from Erciyes Mountain, Turkey. Examination of P. willmanni was conducted with traditional methods [16]. Identification of samples was done by morphologically. Alcohol (75% ethanol) conserved individual specimens of P. willmanni were used for DNA extraction methods. 2.2. DNA Extraction Methods The collected samples were killed in 95% ethanol. For each DNA extraction method, DNA was isolated from individual mites. The sampled DNA was quantified by taking the optical density (OD) measurements at 260 and 280 with a spectrophotometer (ACTGene Micro-Spectrophotometer) and the purity was evaluated by the ratio of OD 260 /OD 280 . The A 260 /A 280 ratio demonstrate the DNA purity, 1.8-2.0 values suggest “pure DNA” [17]. Abstract: We compared three DNA extraction methods; a Chelex resin (C100), Qiagen DNA extraction kit and Cethyl Trimethyl Ammonium Bromide (CTAB) protocol that obtained from Parachipteria willmanni van der Hammen, 1952 mite specimens preserved in ethanol. The aim of the study is to find the most efficient protocol for obtaining DNA from Parachipteria willmanni. All methods are successful in isolating DNA from P. willmanni, but among these three methods, maximum amount of DNA is obtained with Chelex-method. DNAs obtained by these three methods were successfully applied to RAPD-PCR. Keywords: Chelex-method; DNA extraction; Oribatida; Parachipteria willmanni; RAPD-PCR.