Publicação
Identificação de marcadores SSR e de SNPs em medronheiro (Arbutus unedo L.) por sequenciação massiva paralela
| Resumo: | The strawberry tree (Arbutus unedo L.) is native to the Mediterranean region. The use of molecular markers in this species has been limited to the use of RAPDs, ISSRs as well to the cross-amplification of SSRs from other Ericaceae. In this work, we developed a protocol for extracting nuclear DNA from the strawberry tree and performed partial next-generation sequencing of the Arbutus unedo L. genome using the "Ion Torrent" (Life Technologies) platform. The next-generation sequencing resulted in 198,856 sequences ("raw data") with an average size of 123 bp, which were uploaded to the NCBI database "Sequence Read Archive" (SRA) with the accession number: SRX341237. Data analysis led to the identification of 1085 microsatellite-containing sequences, which were also uploaded with accession numbers: from KF023636 to KF024720 to the NCBI databases. Primers were designed for 18 microsatellite loci of which only three have proved to be polymorphic in a panel of 16 samples. Based on identified 25 SNPs one CAPS marker was developed, which despite being heterozygous revealed to be monomorphic among the 16 analyzed samples. |
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| Autores principais: | Fazenda, Pedro Miguel Jesus |
| Assunto: | Arbutus unedo CAPS molecular markers next-generation sequencing nuclear DNA extraction SSRs |
| Ano: | 2013 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | português |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | The strawberry tree (Arbutus unedo L.) is native to the Mediterranean region. The use of molecular markers in this species has been limited to the use of RAPDs, ISSRs as well to the cross-amplification of SSRs from other Ericaceae. In this work, we developed a protocol for extracting nuclear DNA from the strawberry tree and performed partial next-generation sequencing of the Arbutus unedo L. genome using the "Ion Torrent" (Life Technologies) platform. The next-generation sequencing resulted in 198,856 sequences ("raw data") with an average size of 123 bp, which were uploaded to the NCBI database "Sequence Read Archive" (SRA) with the accession number: SRX341237. Data analysis led to the identification of 1085 microsatellite-containing sequences, which were also uploaded with accession numbers: from KF023636 to KF024720 to the NCBI databases. Primers were designed for 18 microsatellite loci of which only three have proved to be polymorphic in a panel of 16 samples. Based on identified 25 SNPs one CAPS marker was developed, which despite being heterozygous revealed to be monomorphic among the 16 analyzed samples. |
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