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lecture_notes:04-06-2015 [2015/04/10 05:44] gepoliano |
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//ILLUMINA SEQUENCING// | //ILLUMINA SEQUENCING// | ||
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Different 3’ and 5’ end adapters – fragments are flanked by the adapters | Different 3’ and 5’ end adapters – fragments are flanked by the adapters | ||
Hybridization in the flowcell (array) | Hybridization in the flowcell (array) | ||
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A washing step takes out one of the two types | A washing step takes out one of the two types | ||
Same cluster: same sequence, sequencing primer | Same cluster: same sequence, sequencing primer | ||
- | Four nucleotides are labeled, all 4 in the same reaction, different 4 calors for each nucleotide | + | Four nucleotides are labeled, all 4 in the same reaction, different 4 colors for each nucleotide |
- | Incorporation by polymerase – light release with colors | + | Incorporation by polymerase – light release with colors. |
- | Same clusters – signal | + | Same clusters – signal. |
CDC camera catches the color. | CDC camera catches the color. | ||
The process continues until ~100 bp | The process continues until ~100 bp | ||
$1000 for a flowcell MinION – 400bp 1 lane | $1000 for a flowcell MinION – 400bp 1 lane | ||
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//PACBIO// | //PACBIO// | ||
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Imagine a plate with small wells. In this technology, the objective is to make a polymerase stick to the well (Eid et al., 2009). This can be imagined as something like ELISA but with only one exactly DNA and polymerase per well. PACBIO is a Real Time sequencing technology, allowing the time it takes to incorporate the base, a measure to infer information about heterochromatin and quadruplex structures. This technology allows long reads. Howver indels are the main mismatch that happens in PacBio. | Imagine a plate with small wells. In this technology, the objective is to make a polymerase stick to the well (Eid et al., 2009). This can be imagined as something like ELISA but with only one exactly DNA and polymerase per well. PACBIO is a Real Time sequencing technology, allowing the time it takes to incorporate the base, a measure to infer information about heterochromatin and quadruplex structures. This technology allows long reads. Howver indels are the main mismatch that happens in PacBio. | ||