scholarly journals Seleção de primers polimórficos para estudo de diversidade genética em cactáceas

2020 ◽  
Vol 2 (3) ◽  
pp. 9
Author(s):  
Daniel Oliveira Jordão do Amaral ◽  
Daniel Rodrigo Cavalcante De Araújo ◽  
Juliana Gomes Freitas ◽  
Fabiane Rabelo da Costa Batista
Keyword(s):  
Polymerase Chain Reaction ◽  
Simple Sequence Repeats ◽  
Chain Reaction ◽  
Pcr Polymerase Chain Reaction ◽  
Inter Simple Sequence Repeats ◽  
Polymerase Chain ◽  
Simple Sequence

A família Cactaceae está distribuída principalmente nas Américas, apresentam uma grande importância econômica, fornecendo recursos energéticos para animais polinizadores e dispersores, podendo ser utilizadas na alimentação animal e humana, possui um grande potencial na medicina tradicional e no paisagismo. O objetivo do presente estudo foi selecionar indicadores e padronizar reações de PCR (Polymerase Chain Reaction) para analisar ISSR (Inter Simple Sequence Repeats) em estudos de variabilidade genética de Cactaceae. Foram testados 14 indicadores de ISSR com temperatura variando de 48° a 52°C, em espécies de Tacinga, e destes, 8 foram selecionados por serem polimórficos: ISSR-808, ISSR-827, ISSR-842, ISSR-845, ISSR-853, ISSR-857 ISSR-880 e ISSR-888. O número médio de sequências amplificadas por indicador foi de 11,5 bandas, com destaque para o indicador ISSR-827, que produziu 15 bandas, enquanto os indicadores ISSR-845 e ISSR-853 produziram apenas 8 bandas. Os 8 indicadores selecionados no presente estudo possibilitaram a diferenciação genética, sendo eficientes e indicando um bom nível de polimorfismo entre as espécies analisadas, dessa forma, poderão ser utilizados em futuros trabalhos para estimar a divergência genética em nível molecular em espécies da família Cactaceae.

Simple sequence repeat primers used in polymerase chain reaction amplifications to study genetic diversity in barley

Genome ◽  
1996 ◽  
Vol 39 (1) ◽  
pp. 112-117 ◽  
Author(s):  
M. P. Sánchez de la Hoz ◽  
J. A. Dávila ◽  
Y. Loarce ◽  
E. Ferrer
Keyword(s):  
Genetic Diversity ◽  
Polymerase Chain Reaction ◽  
Simple Sequence Repeats ◽  
Genetic Relationships ◽  
Arbitrary Sequence ◽  
Chain Reaction ◽  
Barley Cultivars ◽  
Dinucleotide Motif ◽  
Polymerase Chain ◽  
Simple Sequence

In combination with oligonucleotides of arbitrary sequence, 5′ anchored oligonucleotides based on simple sequence repeats were used in polymerase chain reaction amplifications to produce barley DNA fingerprints. The aim of this work was (i) to develop a simple nonradioactive experimental procedure to reveal polymorphism in regions containing SSRs, (ii) to determine the genetic nature of polymorphisms, and (iii) to investigate the efficacy of polymorphisms contained in such fingerprints in disclosing genetic relationships between 14 European barley cultivars with known pedigrees. Different 10-mer oligonucleotides containing a dinucleotide motif were used as single primers and also in pairs with 10-mer oligonucleotides of arbitrary sequence. Further, the arbitrary oligonucleotides were used as single primers to produce RAPDs. Thirteen combinations of primers containing either GT(CA)4 or GC(CA)4 were selected on the basis of number and intensity of scorable bands in silver-stained 7% polyacrylamide gels. Of the fragments scored, 58.4% were polymorphic. Inheritance of these random amplified microsatellite polymorphic fragments (RAMP) was studied in doubled-haploid lines from the F1 of 'Steptoe' × 'Morex'. Fifty percent of the primers generated codominant markers. Genetic similarities between cultivars were estimated from RAMP and RAPD data. Principal coordinate analysis performed on RAMP data revealed a clear separation of winter six-rowed, winter two-rowed, and spring two-rowed barley. The dendograms generated faithfully reflected the genealogies of the barley cultivars. RAPD failed to show clearly the germplasm sources of the experimental cultivars. Key words : simple sequence repeats, microsatellites, amplification, genetic diversity, barley.

Mapping maize microsatellites and polymerase chain reaction confirmation of the targeted repeats using a CT primer

Genome ◽  
1993 ◽  
Vol 36 (5) ◽  
pp. 884-889 ◽  
Author(s):  
M. Lynn Senior ◽  
Manfred Heun
Keyword(s):  
Polymerase Chain Reaction ◽  
Simple Sequence Repeats ◽  
Genome Mapping ◽  
Pcr Amplification ◽  
Chain Reaction ◽  
Expected Size Range ◽  
Pcr Products ◽  
Mammalian Genomes ◽  
Polymerase Chain ◽  
Simple Sequence

Microsatellites, also called simple sequence repeats (SSRs), have yielded an important class of DNA markers most notable for mapping mammalian genomes. To study the occurrence of microsatellites and their inheritance in maize, a search was made of 280 maize GenBank® sequences. Six SSRs were chosen and unique flanking primers were designed for polymerase chain reaction (PCR) amplification. Eight different maize inbreds were studied with these six primer pairs and a mean of 3.5 polymorphic patterns occurred within the expected size range. For five of these putative microsatellites, the segregation in a maize restriction fragment length polymorphism mapping population was analyzed. Four of the microsatellites cosegregated with the Adh1, Gpc1, Pdk1, and Tpi genes from which the primer sequences were derived. The fifth primer pair (MZEGPA1) showed segregating polymorphisms, but the products were larger than expected. To verify the existence of the original SSRs in the segregating PCR products, a CT primer, containing a CT SSR and an arbitrary leader sequence, was used to reamplify these products. The four microsatellites that cosegregated with the original gene were reamplified as anticipated, whereas a suspicious 230-bp product obtained when using the MZEGPA1 primers could not be reamplified. Based on these results it is concluded that microsatellites can be a valuable tool for maize mapping.Key words: maize, microsatellites, simple sequence repeats, genome mapping.

scholarly journals Deleções do DNA Mitocondrial no Envelhecimento: Efeito da Disfunção na Fosforilação Oxida tiva

1999 ◽  
Vol 5 (3) ◽  
pp. 65-66
Author(s):  
Celia Harumi Tengan
Keyword(s):  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Pcr Polymerase Chain Reaction ◽  
Polymerase Chain

Vários estudos descreveram o acúmulo de uma deleção do DNA mitocondrial (DNAmt), denominada de deleção comum, em tecidos pós-mitóticos durante o processo de envelhecimento. Esses achados levaram A hipótese de que radicais livres, gerados dentro da mitocandria, poderiam lesar o DNAmt durante a vida normal. Acredita-se que um defeito no funciomento da cadeia respirat6ra, decorrente da lesão do DNAmt, levaria a um aumento na produção de radicais livres, que por sua vez, lesariam o DNAmt, criando um ciclo vicioso é um fator importante no acúmulo de deleções do DNAmt, pacientes com deficiência da função oxidativa (independente do defeito primário) deveriam apresentar um acúmulo acelerado de deleções do DNAmt. Nós testamos esta hipótese através de três analises: (a) comparação dos níveis de deleção comum em controles normais e pacientes com doenças mitocondriais geneticamente caracterizadas e associadas com uma mutação do DNAmt; (b) análise da cosegregação da deleção comum (associada com o envelhecimento) com uma mutação de ponto patogênica do DNAmt; e (c) detecção de deleções múltiplas do DNAmt através de PCR (polymerase chain reaction) longo em controles e pacientes com doenças mitocondriais. Observamos uma correlação positiva entre a idade e MN/6s de deleção comum em controles (r = 0,80) e pacientes (r = 0,69). As inclinações das curvas eram semelhantes, sugerindo que a taxa de acúmulo da deleção comum associada com a idade era a mesma em ambos os grupos. Não conseguimos observar a co-segregação das moléculas de DNAmt contendo a mutação de ponto com a deleção comum e nem aumento no número de deleções em pacientes. Nossos resultados não suportam a hipótese de que o ciclo vicioso (lesão do DNAmt afeta a função da cadeia respiratória, levando a uma maior produção de radicais livres que, por sua vez, provocaria mais lesão do DNAmt) é um fator importante no acúmulo de deleções do DNAmt no processo de envelhecimento.

Diagnostic biologique précoce de la leptospirose par PCR (polymerase chain reaction)

1993 ◽  
Vol 14 (6) ◽  
pp. 578
Author(s):  
B. Dell'isola ◽  
I. Saint Girons ◽  
P. Amouriaux ◽  
G. Baranton ◽  
A.M. El Maleh ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Pcr Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Diagnostic Biologique

scholarly journals OPTIMALISASI PCR IKAN TONGKOL KRAI (Auxis thazard) DAN LISONG (Auxis rochei) PADA ANALISIS KERAGAMAN GENETIK

2019 ◽  
Vol 11 (2) ◽  
pp. 95
Author(s):  
Raymon Rahmanov Zedta ◽  
Bram Setyadji
Keyword(s):  
Genetic Diversity ◽  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Temperature Range ◽  
Pcr Polymerase Chain Reaction ◽  
Fisheries Resources ◽  
Pcr Product ◽  
Tuna Fishing ◽  
Polymerase Chain

Ikan tongkol lisong dan krai merupakan salah satu jenis tuna yang berperan nyata untuk usaha perikanan tangkap di Indonesia. Pengelolaan sumberdaya ikan tersebut harus selalu dapat dilakukan untuk menjaga tingkat pemanfaatannya supaya tidak lebih tangkap. Kajian keragaman genetik merupakan salah satu teknik dalam pengelolaan pemanfaatan sumberdaya perikanan dengan cara mengetahui tingkat keragaman genetik pada suatu struktur populasi. Kajian keragaman genetik ini diharapkan dapat menjadi basis kajian stok dan opsi dalam pengelolaan sumberdaya perikanan tongkol agar pemanfaatannya dapat dilakukan secara berkelanjutan. Awal mula analisis keragaman genetik dilakukan dengan memperbanyak DNA secara in vitro menggunakan teknik PCR (Polymerase Chain Reaction). Keberhasilan proses PCR dipengaruhi oleh beberapa faktor seperti suhu dan waktu penempelan oligonukleotida primer. Berdasarkan hal tersebut, penelitian ini bertujuan untuk mengetahui suhu dan waktu optimal pada primer Aro2-38. Sampel penelitian diperoleh dari hasil tangkapan pukat cincin yang didaratkan di PPN Palabuhanratu, Jawa Barat. Optimasi PCR menggunakan 12 suhu dan 2 waktu penempelan yang berbeda yaitu : 520C; 52,80C; 540C; 55,50C; 57,20C; 59,10C; 60,90C; 62,80C; 64,50C; 65,90C; 67,20C dan 680C, dan suhu penempelan 30 dan 15 detik. Hasil analisis menunjukkan bahwa produk PCR optimum (menghasilkan pita alel DNA) pada ikan tongkol krai berhasil waktu penempelan 30 detik dengan rentang suhu 52-540C. Sedangkan pada sampel ikan tongkol lisong, produk PCR yang optimum muncul pada waktu penempelan 15 dan 30 detik, dengan rentang suhu 52-60,90C.Frigate and bullet tuna constitute one of tuna species that plays a significant role in Indonesian fishing business. Management of fisheries resources must always be done to maintain the level of utilization so that it is not excessive. Genetic study is one of techniques in managing fisheries resource utilization by knowing the level of genetic diversity in a population structure. This genetic diversity study is expected to be the basis and option in the management of tuna fishing resources so that their utilization can be carried out sustainably. Genetic diversity analysis is start by multiplying fish DNA using PCR (Polymerase Chain Reaction) technique. The success of the PCR process is influenced by several factors such as temperature and time of primary oligonucleotide attachment. Based on this, this study aims to determine the optimal temperature and time in primers Aro2-38. The research sample was obtained from the catch of purse seine landed in PPN Palabuhanratu, West Java. PCR optimization uses 12 temperatures and 2 different annealing times: 520C; 52.80C; 54ÚC; 55,50C; 57.20C; 59.10C; 60.90C; 62.80C; 64,50C; 65,90C; 67.20C and 680C, and the annealing times are 30 and 15 seconds. The results of the analysis showed that the optimum PCR product (producing DNA allele bands) on the cretaceous tuna was successfully pasted for 30 seconds with a temperature range of 52-540C. Whereas in the sample of tuna lisong, the optimum PCR product appeared at the time of attachment of 15 and 30 seconds, with a temperature range of 52-60.90C.

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