Jordans Notes 12 Flashcards

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1
Q

How does PacBio SMRT sequencing work?

A

Sequence by synthesis single strand per well, emitting light from fluorescently labeled nucleotides

Fluorescent tags removed after detection

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2
Q

How inaccurate are PacBio and MinIon?

A

~15% inaccuracy

2nd gen are highly accurate (99%)

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3
Q

What kind of PCR is done with Ion Torrent?

A

Emulsion PCR of fragments bound to microbeads

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4
Q

What kind of PCR is done with Illumina?

A

Solid phase bridge PCR on fragments bound to flow cell

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5
Q

What are the pros and cons of illumina?

A

+
Largest amount of Data
very accurate
-
short reads
Takes a long time
expensive

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6
Q

What are the pros and cons of Ion Torrent?

A

Pros:
- Cheaper
- quicker
- longer read lengths than Illumina

Cons:
- trouble reading homopolymer stretches

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7
Q

What are the pros and cons of PacBio?

A

Pros
-real time sequencing
-long reads
Cons
-Relatively inaccurate

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8
Q

What are the cons of high throughput methods?

A

Huge amounts of data
-processing and storage

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9
Q

What looks at multiple different regions of the
genome at once?

A

Genotyping!

Unlike sequencing which looks at one region at a time

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10
Q

What is the first step of microsatellite genotyping?

A

Finding short tandem repeats and designing primers to target them

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