Next – Generation Sequencing Era Illumina Genome Analyzer Daniel Onofre Vidal, PhD Laboratório de...
Transcript of Next – Generation Sequencing Era Illumina Genome Analyzer Daniel Onofre Vidal, PhD Laboratório de...
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Next – Generation Sequencing Era
Illumina Genome Analyzer
Daniel Onofre Vidal, PhDLaboratório de Genética Molecular e Bioinformática
Curso de Verão - 2010
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Lâmina – 8 canais
1 a 12 amostra por canal = 96 amostras
~375 Mb por canal
3 a 3,5 GB (3 bilhões de bases)
48 horas
Illumina Genome Analyzer
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Illumina Genome Analyzer Workflow
Preparação das bibliotecas
Formação dos clusters
sequenciamento
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Preparação das bibliotecas (6 horas)
Illumina Genome Analyzer
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AA
Preparação das bibliotecas
Illumina Genome Analyzer
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AA
T
T
AAT
T
Preparação das bibliotecas
Illumina Genome Analyzer
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Preparação dos clusters (5 horas)
Illumina Genome Analyzer
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Preparação dos clusters
Illumina Genome Analyzer
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Preparação dos clusters
Illumina Genome Analyzer
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Preparação dos clusters
Illumina Genome Analyzer
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Preparação dos clusters
Illumina Genome Analyzer
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Preparação dos clusters
Illumina Genome Analyzer
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Preparação dos clusters
Illumina Genome Analyzer
A
C
T
G
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Sequenciamento (2-3 dias)
- 4 nucleotídeos marcados ao mesmo tempo- acurácia- sem problemas com homopolimeros
Illumina Genome Analyzer
Incorporaçãodetecçãodesbloqueio
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Sequenciamento
Illumina Genome Analyzer
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Sequenciamento
Illumina Genome Analyzer
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Sequenciamento
Illumina Genome Analyzer
A
TC
G
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Sequenciamento
Illumina Genome Analyzer
AT
C
G
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Sequenciamento
Illumina Genome Analyzer
A
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Sequenciamento
Illumina Genome Analyzer
AT
C
G
A
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Sequenciamento
Illumina Genome Analyzer
A
T
C
G
A
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Sequenciamento
Illumina Genome Analyzer
A
T
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Illumina Genome Analyzer
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Illumina Genome Analyzer
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Illumina Genome Analyzer
Variante não conhecida
SNP
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Illumina Genome Analyzer
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Illumina Genome Analyzer
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Illumina Genome Analyzer
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Illumina Genome Analyzer
Aplicações
Sequenciamento
- DNA (de novo, re-sequenciamento)
- transcriptoma (mRNA seq, tag, small RNA)
Genotipagem ou descoberta
- SNPs
- Copy number variation (CNVs)
- variações estruturais
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Illumina Genome Analyzer
Aplicações
Regulação Gênica
- small RNAs (miRNAs)
Análise Epigenética
- sequencing-based methylation
- DNA-Protein interaction (CHIP-Seq)
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Illumina Genome Analyzer
Aplicações
Regulação Gênica
- small RNAs (miRNAs)
Análise Epigenética
- sequencing-based methylation
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Illumina Genome Analyzer
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Illumina Genome Analyzer
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Illumina Genome Analyzer
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Illumina Genome Analyzer
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Illumina Genome Analyzer
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Illumina Genome Analyzer
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Illumina Genome Analyzer
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Illumina Genome Analyzer
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Illumina Genome Analyzer
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Illumina Genome Analyzer
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Next-Generation Sequencing
Para onde estamos caminhando?
sequenciamento genomas individuais
- 1 a 2 vezes na magnetude do custo
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Obrigado!!!!