Exam 1- Lecture 7

Question 1

What enzymes are used for PacBio SMRT

Answer 1

bound by DNA polymerase
incorporates labelled deoxynucleoside triphosphates

Question 2

What enzymes are used for Oxford Nanopore

Answer 2

sequencing adapter loaded with motor protein

Question 3

What is zero mode waveguide

Answer 3

Nanophotonic devices that confine light to a small observation volume
used in SMRT sequencing

Question 4

How does Oxford Nanopore detect?

Answer 4

DNA attached to sequencing adapter
DNA mixture loaded into flow cell
motor protein unwinds DNA into nanopores
disruptions in current measured and sequence is determined from disruptions in current

Question 5

How does PacBio detect sequences?

Answer 5

SMRTbell, double stranded DNA insert with single stranded hair pin adapters
SMRTbell bound by DNA polymerase and loaded into SMRT Cell with zero-mode waveguides
polymerase processes around SMRTbell template and incorporated fluorescently labelled deoxy nucleoside triphosphate
laser excites fluorophore and then is cleaved
excited fluorophore reveals base identity

Question 6

Is PacBio sequencing by synthesis?

Answer 6

Yes

Question 7

What is the maximum length of DNA per read for each technology?

Answer 7

SMRT- tens of kilobases long
Oxford- 2.3 Mb