Protocolo da extração de DNA de material parafinado para análise de microssatélites em leiomioma

Nascimento, Ericson Martins; Spinelli, Marilda Osti; Rodrigues, Consuelo Junqueira; Bozzini, Nilo

doi:10.1590/s1676-24442003000300013

Cited by 11 publications

(8 citation statements)

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“…The samples that were embedded in paraffin were deparaffinized and rehydrated prior to the steps of DNA extraction protocol, described by Nascimento et al 13 .…”

Section: Dna Extractionmentioning

confidence: 99%

Association of mesenchymal stem cells with platelet rich plasma on the repair of critical calvarial defects in mice

Monteiro

Carlo²,

Neto

et al. 2012

Acta Cir. Bras.

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PURPOSE: To evaluate the effects of mesenchymal stem cells (MSC) from eight mice C57BL/6 gfp+ bone marrows expanded in cultures associated with platelets rich plasma (PRP) deriving from another eight mice, in the repair of critical defects in calvarial bone produced in twenty-four adult isogenic mice C57BL/6. METHODS: The animals were submitted to a cranial defect of 6.0mm in diameter and divided into two equal experimental groups. Control group did not receive treatment and the treated group received a MSC pellet containing 1.0 x 10(7) cells/mL associated with 50.0µL of plasma gel containing 1.0 x 10(9) autologous platelets within the defect. RESULTS: In the treated group was observed process of angiogenesis and bone repair better than control group. CONCLUSION: Mesenchymal stem cells derived from bone marrow of C57BL/6 gfp+ mice associated with PRP gel applied in bone critical defects produced in calvarial contributes positively to the process of bone repair.

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“…The samples that were embedded in paraffin were deparaffinized and rehydrated prior to the steps of DNA extraction protocol, described by Nascimento et al 13 .…”

Section: Dna Extractionmentioning

confidence: 99%

Association of mesenchymal stem cells with platelet rich plasma on the repair of critical calvarial defects in mice

Monteiro

Carlo²,

Neto

et al. 2012

Acta Cir. Bras.

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“…Existem na literatura vários protocolos que descrevem o método de digestão celular. Há autores que relatam a utilização de fervura, acompanhada ou não de solução de lise, e/ou digestão com proteinase K e ainda a utilização desses métodos associados a diferentes tampões (27,38,43) . Outros descrevem a utilização do detergente Triton X-100 e do tiocianato de guanidina para verificar a efetividade dessas substâncias na ruptura ou lise das membranas celulares (14,24) .…”

Section: Digestãounclassified

“…Em muitos desses trabalhos, o kit comercial apresentou DNA de boa qualidade e resultados satisfatórios de amplificação (6,8,13,29,38,39) . Estes são outros kits que em trabalhos científicos mostram ser eficientes na extração DNA de matérias parafinadas: Wizard Genomic DNA Purifications Kit (Promega), Kit GFXtm Genomic Blood DNA Purification, Quick-Gene DNA Tissue Kit S (Fujifilm Life Science), QIAquick Gel Extraction Kit e Kit Glass Max (Gibco BRL, USA) (11,27,34,39) . Uma cuidadosa comparação entre métodos comerciais e in-house em espécimes arquivados foi demonstrada no trabalho de Okello et al (28) .…”

Section: Purificaçãounclassified

Fatores que interferem na qualidade do DNA extraído de amostras biológicas armazenadas em blocos de parafina

Scorsato¹,

Telles²

2011

J. Bras. Patol. Med. Lab.

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Os avanços tecnológicos e metodológicos da biologia molecular criaram a possibilidade de obter DNA de amostras teciduais estocadas em blocos de parafina, alternativa esta que permite estudos retrospectivos de grandes bancos de tecidos contendo inúmeras doenças raras. O objetivo deste trabalho foi a realização de uma breve revisão de alguns fatores inerentes à qualidade do produto extraído proveniente de blocos de parafina armazenados. Os processos envolvidos nas fases de prefixação, fixação, pós-fixação apresentam aspectos que são causa de resultados de produtos finais insatisfatórios. Na fase de prefixação, alterações bioquímicas influenciam na preservação das macromoléculas. As mudanças moleculares causadas pelos fixadores para evitar autólise celular podem ser um limitante no momento da extração de DNA. A qualidade do DNA obtido nas duas primeiras fases é considerada importante para os procedimentos de pós-fixação (extração e purificação do DNA). Existem na literatura vários protocolos, com diferentes passos que podem ser modificados, para a obtenção de DNA de material parafinado. Assim, a compreensão das reações em cada fase é importante para solucionar ou minimizar problemas, que influenciam na qualidade das macromoléculas de DNA.

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“…Furthermore, the archiving time of paraffined parts can make the DNA more susceptible to degradation. Thus, it is essential to set up appropriate extraction methods that make it possible to recover DNA from this type of material [3,4].…”

Section: Introductionmentioning

confidence: 99%

DNA extraction and purification from archival material: Revisiting the standard operating procedure

Netto¹,

Alves²,

Carvalho

et al. 2015

Forensic Science International: Genetics Supplement Series

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A B S T R A C TForensic evidence from tissues embedded in paraffin archived for long periods and safeguarded in Pathology Institutes are, in some cases, the only sample available for DNA analysis. However, recovering DNA from this material can be challenging. The aim of this study is to evaluate methods of extraction and purification of DNA, using biological archival materials, in order to determine a protocol that is easy use and efficiency to adapt to the Institute of Research and Expertise in Forensic Genetics (IPPGF). Twentytwo samples from cadaver tissues included in paraffin blocks were submitted to two pre-treatments: deparaffinization using xylene and heating in microwave. After that, four methods were used for DNA extraction: phenol-chloroform-isoamyl alcohol (PCI), Chelex 100 1 , Purification on membranes (NucleoSpin) and the Resin purification method (DNA IQ). The quality and quantity of the DNA extracted was compared using RT-PCR. Additionally, PCR reactions were performed using the MiniFiler TM Kit. The methodology that presented better DNA recovery and more loci amplified in relation to the age of the tissues and also for the different tissues embedded in paraffin was PCI. These results reinforce and validate the maintenance of Standard Operating Procedure based on PCI used in IPPGF.

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Protocolo da extração de DNA de material parafinado para análise de microssatélites em leiomioma

Cited by 11 publications

References 0 publications

Association of mesenchymal stem cells with platelet rich plasma on the repair of critical calvarial defects in mice

Association of mesenchymal stem cells with platelet rich plasma on the repair of critical calvarial defects in mice

Fatores que interferem na qualidade do DNA extraído de amostras biológicas armazenadas em blocos de parafina

DNA extraction and purification from archival material: Revisiting the standard operating procedure

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