PacBio Flashcards
1
Q
What can PacBio sequence?
A
RNA and DNA from any sample type
2
Q
What are the steps in PacBio?
A
- DNA/RNA isolated
- Ligate adapters to dsDNA (creating circular template)
- Primer and polymerase added to library.
- Library placed on instrument for sequencing.
3
Q
What do SMRT Cells contain?
A
Contains millions of small wells called Zero-mode wave guides (ZMW).
4
Q
What is in the Zero-Mode Wave guides (ZMW)?
A
A single immobilised DNA molecule.
5
Q
What is emitted during nucleotide incorporation?
A
Light
6
Q
There are two sequencing modes, what is Circular Consensus Sequencing used for?
A
highly accurate long reads (called Hifi reads)
7
Q
There are two sequencing modes, what is Continuous Long Read Sequencing used for?
A
To generate the longest possible read.
8
Q
What powers PacBio?
A
by Single Molecule Real-Time (SMRT) Sequencing
