DNA extraction in mangaba tree (Hancornia speciosa Gomes)

Nascimento, Ana Letícia Sirqueira; Sá, Aline Jesus; Lédo, A. da S.; Silva, Ana Veruska Cruz da

doi:10.3738/1982.2278.2727

Cited by 4 publications

(4 citation statements)

References 1 publication

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“…Based on this result, the DNA of each strain (5, 5SF, and 5CF) was then individually extracted with DNAzol. DNA extraction methods usually vary according to the sample or matrix (Nascimento et al, 2017). However, the solutions must contain a buffer substance to stabilize the pH, a salt to dissociate the proteins, a detergent to solubilize the membranes, and an agent to inactivate the DNases and protect the genomic DNA (Nascimento et al, 2017).…”

Section: Resultsmentioning

confidence: 99%

Isolation, molecular identification, and in vitro control of Campomanesia rufa (O. Berg) Nied. endogenous bacteria

Mosqueira,

Paiva,

Rivas

et al. 2023

Ciênc. agrotec.

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Campomanesia rufa (O. Berg) Nied. is a Myrtaceae species native to the Brazilian Cerrado. As C. rufa is considered endangered, in vitro propagation is an alternative for its conservation. However, the lack of effective disinfection protocols for endophytic microorganisms naturally present in plant tissues hinders the success of micropropagation. In this context, the objective of this study was to isolate, molecular identify, and control endogenous occurring bacteria of C. rufa propagated in vitro. Purified PCR products of bacterial isolates were sequenced by the Sanger method and aligned with homologous sequences using the Basic Local Alignment Search Tool (BLAST) available in National Centre for Biotechnology Information (NCBI) database. MEGA7 software was used to align all sequences and to draw phylogenetic trees. Survival, shoot height, and efficiency of antibiotics (streptomycin, ampicillin, and chloramphenicol) at different concentrations in the culture medium were evaluated. Different Bacillus sp. strains were observed in the plant tissues. When testing the control of Bacillus sp. with antibiotics, 32 mg L-1 ampicillin caused a significant reduction in bacterial contamination with no effect on explant survival. On the other hand, 256 mg L-1 streptomycin caused the greatest reduction in contamination but was lethal to over 90% of the explants. This study is the first report on the occurrence of endogenous bacteria and their control in the context of in vitro native species conservation.

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Section: Resultsmentioning

confidence: 99%

Isolation, molecular identification, and in vitro control of Campomanesia rufa (O. Berg) Nied. endogenous bacteria

Mosqueira,

Paiva,

Rivas

et al. 2023

Ciênc. agrotec.

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“…Regarding the ISSR response in the PCR reaction, the annealing temperature is known to be a factor that directly contributes to the specificity of the molecular marker. In this context, the ISSR marker has a varied annealing temperature, ranging from 40 to 55 °C (NASCIMENTO et al, 2017), when compared to other dominant markers, such as RAPD (Random Amplified Polymorphic DNA), which anneal at a temperature between 34 and 37 °C (MISHRA et al, 2015;PRIYA;MANIMEKALAI;RAVICHANDRAN, 2015).…”

Section: Resultsmentioning

confidence: 99%

AMPLIFICATION AND SELECTION PROFILE OF ISSR MARKERS FOR GENETIC STUDIES IN Calotropis procera

et al. 2022

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Sodom apple is a plant species adapted to various ecosystems and has stood out for its economic and ecological importance. We evaluated the amplification profile of 23 ISSR primers and selected polymorphic loci for genetic studies of a natural population of Calotropis procera by collecting and extracting genomic DNA from 33 individuals. Genomic DNA was extracted using the sorbitol protocol and 2% CTAB and the ISSR amplification products were resolved by electrophoresis. Based on the amplification profile, the 23 primers were classified as suitable, moderate, and unsuitable. We described the quality of primers considering the total number of bands, mean bands per primer, percentage of polymorphism, Nei’s genetic diversity (expected heterozygosity – He), assuming Hardy-Weinberg equilibrium and the polymorphic information content (PIC). All ISSR primers showed an amplification profile, which generated 173 bands with an average of 7.5 loci per primer. However, only 18 out of the 23 tested primers allowed visible and high-quality amplification, which were classified as suitable and polymorphic. We also observed a mean of 0.30 and 0.24 for PIC and He estimates, respectively. The DiCA3`RG, TriAGA3`RC, and TriCGC3`RC primers were highly transferable to C. procera (they presented quality for amplification with good reproducibility), with PIC values higher than 0.40, He higher than 0.30, and polymorphism higher than 86%.

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“…Já a contaminação com proteínas é evitada pelo uso de reagentes de precipitação como o cloreto de sódio, modulação da temperatura da reação ou aplicação de enzimas degradadoras (Pereira et al, 2019). Nesse estudo, o uso da proteinase K, embora eleve o custo do método, é considerado um reagente eficaz por promover desproteinização de maneira eficaz e uma alternativa segura comparada ao uso de reagentes de elevada toxicidade como tiocianato de guanidina, fenol e clorofórmio (Nascimento et al, 2017;Qamar et al, 2017;Rodriguez et al, 2017).…”

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Adaptação de um método de extração de DNA a partir de células da mucosa bucal baseado em parâmetros de qualidade / Adaptation of a DNA extraction method from buccal mucosa cells based on quality parameters

Araujo¹,

Valente²,

Souza³

et al. 2021

BJDV

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Umas das técnicas mais utilizadas para a análise de doenças genéticas é a extração de DNA para fins diagnósticos e de pesquisas. E a coleta das células da mucosa bucal por meio de swab se mostra um método ideal, pois possui uma abordagem segura, indolor, e eficiente para obtenção de DNA. Assim, o objetivo deste trabalho foi adaptar um método de extração de DNA a partir de células da mucosa bucal baseado em parâmetros de qualidade. Para isso, foi realizada a coleta da mucosa bucal de dez voluntários com swab para extração de DNA por meio do protocolo adaptado. Os parâmetros avaliados foram à quantidade, integridade e pureza do DNA obtido. Ainda para avaliar a aplicabilidade da técnica na amplificação de genes foi realizada a reação em cadeida da

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DNA extraction in mangaba tree (Hancornia speciosa Gomes)

Cited by 4 publications

References 1 publication

Isolation, molecular identification, and in vitro control of Campomanesia rufa (O. Berg) Nied. endogenous bacteria

Isolation, molecular identification, and in vitro control of Campomanesia rufa (O. Berg) Nied. endogenous bacteria

AMPLIFICATION AND SELECTION PROFILE OF ISSR MARKERS FOR GENETIC STUDIES IN Calotropis procera

Adaptação de um método de extração de DNA a partir de células da mucosa bucal baseado em parâmetros de qualidade / Adaptation of a DNA extraction method from buccal mucosa cells based on quality parameters

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