Developing a rapid, efficient and low cost method for rapid DNA extraction from arthropods

Mega, Nicolás Oliveira; Revers, L. F.

doi:10.1590/s0103-84782011005000112

Cited by 5 publications

(4 citation statements)

References 16 publications

(17 reference statements)

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“…To our knowledge, this is the fastest protocol for extraction of DNA from a single individual of soft-bodied, small insects reported to date. Serval low-cost DNA extraction methods are known where the estimated cost ranges from US$0.1-0.5 [50,51]. Whereas, the present study reports the zero-cost protocol for extraction of insect DNA.…”

Section: Plos Onementioning

confidence: 89%

Rapid and zero-cost DNA extraction from soft-bodied insects for routine PCR-based applications

Jangra

Ghosh

2022

PLoS ONE

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Nucleic acid extraction is the first and foremost step in molecular biology studies. Extraction of DNA from small, soft-bodied insects is often time-consuming and costly. A fast, easy, and cost-effective DNA extraction method with greater yield and purity of DNA would aid in the rapid diagnostics, screening of large populations, and other routine PCR-based applications. The present study evaluated and standardized a rapid and zero-cost DNA extraction from soft-bodied small insects for routine molecular studies. Five rapid DNA extraction methods viz. extraction in sterile distilled water (SDW), 1X phosphate-buffered saline (PBS, pH 7.4), 1.4 M sodium chloride (NaCl), 20 mM ethylenediaminetetraacetic acid (EDTA, pH 8.0), and elution from blotted nitrocellulose membrane (NCM) were compared with standard CTAB extraction buffer and DNeasy® Blood and Tissue Kit methods. The average yield, purity, storage stability, time, and cost of extraction were assessed for all the methods and compared. A method of DNA extraction by simply crushing the soft-bodied insects in SDW was ideal in terms of yield, purity, storability, and performing routine PCR-based applications including detection of pathogens from vector species. The extraction could be accomplished in 2.5 min only with zero-reagent cost. The method would be useful in rapid molecular diagnostics and screening large populations of soft-bodied insects.

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Section: Plos Onementioning

confidence: 89%

Rapid and zero-cost DNA extraction from soft-bodied insects for routine PCR-based applications

Jangra

Ghosh

2022

PLoS ONE

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“…Total DNA for genotyping was extracted from adult individuals following the protocol described by Mega and Revers (2011) and diluted to a concentration of 50 ng/μL for use in AFLP and SSR assays.…”

Section: Methodsmentioning

confidence: 99%

Association between molecular markers and behavioral phenotypes in the immatures of a butterfly

et al. 2018

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Newly hatched caterpillars of the butterfly Heliconius erato phyllis routinely cannibalize eggs. In a manifestation of kin recognition they cannibalize sibling eggs less frequently than unrelated eggs. Previous work has estimated the heritability of kin recognition in H. erato phyllis to lie between 14 and 48%. It has furthermore been shown that the inheritance of kin recognition is compatible with a quantitative model with a threshold. Here we present the results of a preliminary study, in which we tested for associations between behavioral kin recognition phenotypes and AFLP and SSR markers. We implemented two experimental approaches: (1) a cannibalism test using sibling eggs only, which allowed for only two behavioral outcomes (cannibal and non-cannibal), and (2) a cannibalism test using two sibling eggs and one unrelated egg, which allowed four outcomes [cannibal who does not recognize siblings, cannibal who recognizes siblings, “super-cannibal” (cannibal of both eggs), and “super non-cannibal” (does not cannibalize eggs at all)]. Single-marker analyses were performed using χ2 tests and logistic regression with null markers as covariates. Results of the χ2 tests identified 72 associations for experimental design 1 and 73 associations for design 2. Logistic regression analysis of the markers found to be significant in the χ2 test resulted in 20 associations for design 1 and 11 associations for design 2. Experiment 2 identified markers that were more frequently present or absent in cannibals who recognize siblings and super non-cannibals; i.e. in both phenotypes capable of kin recognition.

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“…protokol yang murah, tetapi perlu waktu untuk persiapan reagen dan optimasi untuk menghasilkan DNA yang berkualitas. Kit komersial dapat melakukan ekstraksi DNA dengan cepat, tidak menggunakan kloroform atau fenol yang berbahaya, tetapi harganya lebih mahal jika dibandingkan dengan protokol home made (Mega dan Revers, 2011). Perbandingan hasil ekstraksi DNA menggunakan beberapa protokol dari kit komersial atau protokol home made merupakan kajian menarik yang dapat dilakukan lebih lanjut.…”

Section: Hasil Dan Pembahasanunclassified

OPTIMATION OF DNA EXTRACTION PROTOCOL OF Elaeidobius kamerunicus

Wening

Prasetyo

Rozziansha

et al. 2018

J. Pen. Kelapa Sawit

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African pollination weevil (Elaeidobius kamerunicus Faust) has an important role in the productivity of Indonesian oil palm plantation. Up to now, there has not been a comprehensive biological study of the species at molecular level. The basic knowledge is very useful for exploitation of the weevil for effective oil palm fruit set development. This research aimed to obtain DNA extraction protocol of E. kamerunicus for DNA fingerprinting of the species. Results showed that using a DNA extraction kit,material disruption by using micro pestle resulted the highest quantity of DNA, while there were no significant differences of resulted DNA quantity among treatments using tissue lyser for material disruption. DNA extracted by using micro pestle or tissue lyser for material disruption is adequate for DNA fingerprinting using AFLP (Amplified Fragment Length Polymorphism) and sequencing techniques.

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Developing a rapid, efficient and low cost method for rapid DNA extraction from arthropods

Abstract: Here, it is presented a rapid and efficient method

Cited by 5 publications

References 16 publications

Rapid and zero-cost DNA extraction from soft-bodied insects for routine PCR-based applications

Rapid and zero-cost DNA extraction from soft-bodied insects for routine PCR-based applications

Association between molecular markers and behavioral phenotypes in the immatures of a butterfly

OPTIMATION OF DNA EXTRACTION PROTOCOL OF Elaeidobius kamerunicus

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