Explain PacBio Sequencing

Expert Solution

PacBio offered an alternate sequencing methodology called Single-molecule real-time (SMRT) sequencing. As it is developed by PacBio, it is also called PacBio sequencing. This is a third-generation sequencing technology

advantages :

1. longer read lengths

2. faster runs

How it works:

1. Choose Double stranded DNA sample to be sequenced

2. Hairpin adapaters will be ligated to end of the DNA sample.

3. The double strand DNA sample will now get converted into single stranded circular DNA template called SMRTbell.

4. SMARTbell will be loaded into a chip called SMRT cell. The SMRT cell has a sequencing unit called Zero Mode wave guide. ZMW has single polymerase immobilized at the bottom. The polymerase can bind to adapters and start replications.

5. Fluorescently tagged nucleotides are added in the SMRT cell. Thus upon replication by polymerase, distinct emission spectrum gets generated.

6. The emission helps in identifying the base added to template by Polymerase. Thus sequencing data will be collected and processed.

gladiator answered 1 year ago

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