Nanopore

Question 1

What does Oxford Nanopore use?

Answer 1

pore-forming proteins creating membrane pores

Question 2

How can nanopores gain different shapes and chemical modifications?

Answer 2

Through engineering key residue mutations in the barrel of the pore.

Question 3

How do nanopores help sequence DNA/RNA?

Answer 3

• nanopore creates hole in membrane
• current characteristically disrupted when DNA molecule and nanopore come into contact
• current disruption can be measured
• can interpret analyte through current disruption

Question 4

What components make up nanopore sequencing?

Answer 4

• electronically resistant membrane
• motor protein
• nanopore reader

Question 5

What is the role of a motor protein during nanopore sequencing?

Answer 5

1. Controls the speed in which DNA and RNA pass through the nano pore.
2. Unzips the double strand as it feeds it through the nanopore

Question 6

What speed should the RNA and DNA go through the pore?

Answer 6

DNA: 450 bp/second
RNA: 70 bp/second

Question 7

How does the nanopore reader work?

Answer 7

Contains combination of amino acids.
Amino acids discriminate against nucleotides in the pore.
Reader reads nucleotides and creates a word for every four bases.

Question 8

How is the raw signal processed?

Answer 8

Recurrent Neural Network (RNN)

Question 9

What are nanopores arranged on?

Answer 9

Sensor array

Question 10

What is the Action Specific Integrated Circuit (ASIC)?

Answer 10

Measures ionic current flow through each nanopore in the sensor array chip.

Question 11

How is a current generated through the nanopore?

Answer 11

By applying a potential across the membrane.

(Applying an electrical field between two points)

Question 12

What is a flow cell?

Answer 12

Component of Oxford Nanopore sequencing device.
Consists of sensor array, Application-Specific Integrated Circuit (ASIC), and nanopores.