Comparative analysis of protocols for DNA extraction from soybean caterpillars

Abstract

ABSTRACT. Genomic DNA extraction is crucial for molecular research, including diagnostic and genome characterization of different organisms. The aim of this study was to comparatively analyze protocols of DNA extraction based on cell lysis by sarcosyl, cetyltrimethylammonium bromide, and sodium dodecyl sulfate, and to determine the most efficient method applicable to soybean caterpillars. DNA was extracted from specimens of Chrysodeixis includens and Spodoptera eridania using the aforementioned three methods. DNA quantification was performed using spectrophotometry and high molecular weight DNA ladders. The purity of the extracted DNA was determined by calculating the A260/A280 ratio. Cost and time for each DNA extraction method were estimated and analyzed statistically. The amount of DNA extracted by these three methods was sufficient for PCR amplification. The sarcosyl method yielded DNA of higher purity, because it generated a clearer pellet without viscosity, and yielded high quality amplification products of the COI gene I. The sarcosyl method showed lower cost per extraction and did not differ from the other methods with respect to preparation times. Cell lysis by sarcosyl represents the best method for DNA extraction in terms of yield, quality, and cost effectiveness.

A Simple and Efficient Method of Extracting DNA from Aged Bones and Teeth

“…Due to the special preservation environment of the old skeletal and tooth samples, the release of DNA is facilitated by the digestion and dissolution of the cell membrane by surfactants according to the literature. NLS is an anionic surfactant that is more suitable for DNA extraction than SDS and cetrimonium bromide (CTAB) based on the yield, quality, and cost-effectiveness (8,25). By optimizing the solution composition, concentration, dosage, temperature, and time for digestion, the best results were achieved by adding 250 lL of digestion solution (0.3 M EGTA and 0.5% NLS) to the 50 mg of bone or tooth powder at 56°C for 1 h in the dark.…”

Minimally destructive hDNA extraction method for retrospective genetics of pinned historical Lepidoptera specimens

“…DTT, PTB, or CTAB) for making the DNA recoverable 14 , choosing the most efficient extraction buffer is a crucial step for the extraction of DNA of sufficient quality and quantity from both modern and in particular historical specimens. Although dozens of different extraction protocols have been published, few studies have investigated which one is most efficient for a specific taxonomic group, type of tissue, or preservation type 15 , 16 . Published studies for pinned and historic insects use a variety of protocols, sometimes without providing any additional rationale for the protocol selection beyond the shared use of using historic/ancient material 14 , 17 – 18 , despite indications that DNA recovery from historical material might be enhanced by carefully choosing the extraction buffers 20 .…”

An optimized high-quality DNA isolation protocol for spodoptera frugiperda J. E. smith (Lepidoptera: Noctuidae)

“…For most Hemiptera species, high-yield/-quality DNA isolation is severely affected by a variety of inhibitory compounds found in the DNA extraction matrix, mainly coming from the processed biological sample (e.g., polysaccharides or phenols) or the chemical used for making the DNA accessible (e.g., CTAB) [21] , [22] , [23] . However, few studies deal with a comparison of extraction methods intended to find the most appropriate method to use in each species or family [21 , 24 , 25] . These insects contain plant phenolics and tannins in their digestive tracts, especially when the insects are not adults, and they are not easily dissected for DNA isolation.…”