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19 July 2019 Comparison of five DNA extracting protocols from Typhlodromus (Anthoseius) bagdasarjani Wainsten and Arutunjan (Acari: Phytoseiidae)
Asghar Hosseininia, Mohammad Khanjani, Mahdieh Asadi, Jalal Soltani
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Abstract

Obtaining clean DNA from mites to be used in molecular studies has always been a challenging task. In this study, five protocols (CTAB-based handmade kit, salting out protocol, STE protocol, modified CTAB-based protocol and wizard Genomic DNA purification kit) were evaluated to extract DNA from Typhlodromus (Anthoseius) bagdasarjani. Quantitative and qualitative analyses of the DNA concentrations and a comparison between the ratio of absorption of wavelength from 260 to 280 nm in a completely randomized split plot design for five extraction protocols and three levels of 1, 5, 10 mites were performed. PCR protocol was used to examine the amplification of the ITS1 fragment from the specimens. The results showed that wizard kit and CTAB modified protocol were the bests. However, economically, modified CTAB-based protocol is the best choice for extensive experiments, since this protocol allows extracting DNA from an individual mite and provides the highest concentration DNA (143.35±0.85ng) and the best OD (1.80±0.03) for molecular studies.

© Systematic & Applied Acarology Society
Asghar Hosseininia, Mohammad Khanjani, Mahdieh Asadi, and Jalal Soltani "Comparison of five DNA extracting protocols from Typhlodromus (Anthoseius) bagdasarjani Wainsten and Arutunjan (Acari: Phytoseiidae)," Systematic and Applied Acarology 24(7), 1249-1260, (19 July 2019). https://doi.org/10.11158/saa.24.7.9
Received: 26 March 2019; Accepted: 26 May 2019; Published: 19 July 2019
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KEYWORDS
CTAB
DNA extraction
extraction kit
predatory mite
Typhlodromus (Anthoseius) bagdasarjani
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