Comparative analysis of protocols for DNA extraction from soybean caterpillars

被引:8
作者
Palma, J. [1 ]
Valmorbida, I. [2 ]
da Costa, I. F. D. [3 ]
Guedes, J. V. C. [4 ]
机构
[1] Cooperat Cent Gaucha Ltda, Unidade Tecnol, Setor Entomol, Cruz Alta, RS, Brazil
[2] Univ Fed Santa Maria, Ctr Ciencias Rurais, Santa Maria, RS, Brazil
[3] Univ Fed Santa Maria, Dept Def Fitossanitaria, Ctr Ciencias Rurais, Programa Posgrad Agron,Clin Fitossanitaria, Santa Maria, RS, Brazil
[4] Univ Fed Santa Maria, Dept Def Fitossanitaria, Ctr Ciencias Rurais, Lab Manejo Integrado Pragas,Programa Posgrad Agro, Santa Maria, RS, Brazil
关键词
Chrysodeixis includens; PCR; Spodoptera eridania; Sarcosyl; SPODOPTERA-FRUGIPERDA LEPIDOPTERA; MOLECULAR VARIABILITY; NOCTUIDAE; BRAZIL; MAIZE; PLANT; RAPD;
D O I
10.4238/gmr.15027904
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
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页数:9
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