PCR is another in vitro DNA synthesis reaction The twist in this case is that the reaction is repeated 25-30 times so that the DNA between the primers.

Apresentação em tema: "PCR is another in vitro DNA synthesis reaction The twist in this case is that the reaction is repeated 25-30 times so that the DNA between the primers."— Transcrição da apresentação:

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2 PCR is another in vitro DNA synthesis reaction The twist in this case is that the reaction is repeated 25-30 times so that the DNA between the primers is amplified

3 The first PCR cycle: The sequence between the two primers will be amplified

4 Four cycles of PCR

5 Copy number of the sequence between the primers increases exponentially

6 SEQUENCIAMENTO DE DNA Profa. Dra. Maria Aparecida Fernandez Depto de Biologia Celular e Genética Universidade Estadual de Maringá

7 Structure of dideoxynucleotide triphosphates

8 Dideoxy DNA sequencing is an in vitro DNA synthesis reaction with a twist DNAP requires a template and a primer The [ddNTP] determines the distribution of chain lengths produced. The primer is labeled so the DNA fragments synthesized can be detected by autoradiography.

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10 TGGCTCGGCCTCAAGTCGAG GTTATCAGATCTGCAACTCAA Alto peso molecular Baixo peso molecular Filme de raio X Auto-radiogramaLeitura manual

11 Automated DNA Sequencing

12 Reação de seqüênciamento

13 Fragmentos amplificados na reação de seqüênciamento

14 Typical output of an automated sequencer

15 Eletroforese no seqüênciamento

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17 Captura do fluorescente e processamento da informação

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20 Seqüenciador automático

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29 Conjunto de 16 capilares

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34 Panorama no momento da corrida

35 Análise preliminar pós corrida

36 ELETROFEROGRAMAS