Effects of Different Pretreatments of DNA Extraction from Dried Specimens of Ladybird Beetles (Coleoptera: Coccinellidae).

Weidong Huang,Xiufeng Xie,Xinyue Liang,Xiaosheng Chen,Xingmin Wang

doi:10.3390/insects10040091

Abstract

Obtaining genetic information from museum specimens is a fundamental component of many fields of research, including DNA barcoding, population genetics, conservation genetics, and phylogenetic analysis. However, acquiring genetic information from museum specimens is challenging because of the difficulty in amplifying the target sequences due to DNA damage and degradation. Different pretreatments can significantly impact the purity and concentration of genomic DNA from museum specimens. Here, we assessed four pretreatment methods—use of 0.9% NaCl buffer, phosphate-buffered saline (PBS), Saline Tris-EDTA (STE) buffer, and sterile water—to determine which pretreatment is most suitable for DNA extraction from dried specimens of ladybird beetles. We completed a comprehensive phylogenetic analysis to test whether the sequences obtained from dried specimens enable proper phylogenetic inference. Our results showed that pretreatment can improve the quality of DNA from dried specimens. The pretreatment effects of 0.9% NaCl buffer and STE buffer were better than those of PBS buffer and sterile water. The phylogenetic analyses results showed that museum specimens can be used to generate cogent phylogenetic inferences. We report the optimum pretreatment methods for DNA extraction from dried ladybird beetles specimens as well as provide evidence for accurately determining phylogenetic relationships for museum specimens.

Highlights

Developments in molecular biology, specimen identification, and phylogenetic and population genetics require the use of molecular techniques, and DNA sequences provide vast quantities of information for phylogenetic inference and taxonomic identification
Our results indicate that different pretreatments can improve and concentration, and the 0.9% NaCl buffer and Saline Tris-EDTA (STE) buffer produced the highest DNA purity and quality
We assessed the effects of four pretreatment methods—bathing in 0.9% NaCl buffer, phosphate-buffered saline (PBS) buffer, STE buffer, and sterile water—to identify the best practice for high-quality DNA extractions using dried ladybird beetles specimens

Summary

Introduction

Developments in molecular biology, specimen identification, and phylogenetic and population genetics require the use of molecular techniques, and DNA sequences provide vast quantities of information for phylogenetic inference and taxonomic identification. Obtaining and collecting fresh material is time-consuming, expensive, and often fails to provide a wide coverage of the species [1]. Museum specimens generally cover a broader taxonomic range and are more obtained, enabling a wider range of questions and taxa to be studied [2]. Many researchers are trying to exploit this potential of museum specimens [3,4,5] and some museums are becoming active molecular genetics research institutions. Museums worldwide house millions of animal and plant specimens, many of which have been preserved so that scientific investigation is possible.

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