Jordans Notes 12

Question 1

How does PacBio SMRT sequencing work?

Answer 1

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

Fluorescent tags removed after detection

Question 2

How inaccurate are PacBio and MinIon?

Answer 2

~15% inaccuracy

2nd gen are highly accurate (99%)

Question 3

What kind of PCR is done with Ion Torrent?

Answer 3

Emulsion PCR of fragments bound to microbeads

Question 4

What kind of PCR is done with Illumina?

Answer 4

Solid phase bridge PCR on fragments bound to flow cell

Question 5

What are the pros and cons of illumina?

Answer 5

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

Question 6

What are the pros and cons of Ion Torrent?

Answer 6

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

Cons:
- trouble reading homopolymer stretches

Question 7

What are the pros and cons of PacBio?

Answer 7

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

Question 8

What are the cons of high throughput methods?

Answer 8

Huge amounts of data
-processing and storage

Question 9

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

Answer 9

Genotyping!

Unlike sequencing which looks at one region at a time

Question 10

What is the first step of microsatellite genotyping?

Answer 10

Finding short tandem repeats and designing primers to target them