What is the function of zero-mode waveguide in SMRT sequencing?
The zero-mode waveguide is essentially that micro-microwave door. Within the ZMW, a molecule of DNA polymerase, alongside the template DNA that is to be sequenced, is adsorbed to the bottom of the base, and nucleotides with fluorescent molecules attached are released into the ZMW chamber.
How does the zero-mode waveguide work?
Zero-mode waveguides are photonic nanostructures that create highly confined optical observation volumes, thereby allowing single-molecule-resolved biophysical studies at relatively high concentrations of fluorescent molecules.
In which technique a zero-mode waveguide chamber is used?
Zero-mode waveguides have been developed for rapid parallel sensing of zeptolitre sample volumes, as applied to gene sequencing, by Pacific Biosciences (previously named Nanofluidics, Inc.)
What is the detection volume of a zero-mode waveguide Zmw used in PacBio SMRT )?
10-21 liters
SMRT Technology At-a-Glance Each ZMW becomes a nanophotonic visualization chamber providing a detection volume of just 20 zeptoliters (10-21 liters). At this volume, the activity of a single molecule can be detected amongst a background of thousands of labeled nucleotides.
How does long read sequencing work?
Long-read, or third-generation, sequencing involves reading sequences of between 10,000 and 100,000 base pairs in one go (although much longer reads have also been reported), without the need to cut up and amplify DNA samples.
What is waveguide in optical fiber?
An optical waveguide is a physical structure that guides electromagnetic waves in the optical spectrum. Common types of optical waveguides include optical fiber, transparent dielectric waveguides made of plastic and glass, liquid light guides, and liquid waveguides.
Is a lens a waveguide?
Waveguide lenses use this same property of a unidirectional light-wave to guide a wave of light through a lens or plane (planar waveguide), from one end to the other. Some of the most basic waveguides were first commercialized by Lumus Optics (coming out of Israeli military-industrial research).
What is PacBio sequencing?
PacBio sequencing captures sequence information during the replication process of the target DNA molecule. The template, called a SMRTbell, is a closed, single-stranded circular DNA that is created by ligating hairpin adaptors to both ends of a target double-stranded DNA (dsDNA) molecule (Figure 1) [2].
Why is long-read sequencing better?
Linked-reads allow for a significantly increased physical coverage with only a slight increase of standard sequencing, ultimately providing better access to typically inaccessible parts of the genome and overcoming several of the limitations of short-read sequencing.