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Issue:ISSN 2095-1353
           CN 11-6020/Q
Director:Chinese Academy of Sciences
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Your Position :Home->Past Journals Catalog->2023年60 No.6

Comparison of three methods of extracting DNA from acaroid mites without causing morphological damage to specimens
Author of the article:FAN Hai-Ye, ZHU Yong-Hang, CHEN Hui-Lin, LIU Lu-Yao, ZHAO Wei, CAO Yu-Xiang, YE Chang-Jiang, SUN En-
Author's Workplace:School of Laboratory Medicine, Wannan Medical College
Key Words:acaroid mites; DNA extraction; without morphological damage; modified Chelex-100 method; alkaline lysis method
Abstract:

Abstract  [Objectives]  To evaluate three methods of extracting DNA from acaroid mites without causing morphological damage, thereby preserving complete voucher specimens. [Methods] Genomic DNA of a single Aleurolyphus ovatus was extracted using the modified Chelex-100 method, the STE method, and the Alkaline lysis method, followed by voucher specimen recovery and morphological comparison. The purity of DNA was determined using an ultra microspectrophotometer. The COI gene was amplified to verify whether the extracted DNA met PCR amplification requirements. [Results]  Voucher specimens subjected to one of the 3 methods remained intact as a whole, but the alkaline lysis method caused excessive transparency of the cuticle and detachment of body bristles, compromising the morphological identification of specimens. The A260/A280 ratio of the modified Chelex-100 method was 1.95, whereas that of the other two methods was < 1.8. DNA extracted by either of the three methods met the standard for PCR amplification, although the degree of amplification achieved using the STE method was relatively poor. [Conclusion] The modified Chelex-100 method had the least effect on the morphology of voucher specimens and did not compromise subsequent morphological identification. DNA extracted using this method also has high purity, sufficient to meet the requirements of PCR amplification and subsequent molecular research. The Chelex-100 method is simple and efficient, and allows DNA to be extracted from single acaroid mites without morphological damage.

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