Nanopore sequencing is one of the most advanced techniques for gene/genome sequencing.

1. N A N O P O R E
S E Q U E N C I N G
Kuldeep Gauliya

2. Why Nanopore ??
• Flexibility and versatility
• Single molecule sequencing
• Both short and long read lengths
• Real-time data generation and analysis
• Cost-effectiveness
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3. Introduction
The concept was envisioned in the early 1990s by David Deamer (UC
Santa Cruz) and Daniel Branton (Harvard).
• UC Santa Cruz Nanopore Group researchers invented the idea of
nanopore sequencing.
• Goal of sequencing human genomes for $1000 or less.
• These nanopore instruments are built around a “nano-scale” opening
in a thin membrane, for the study of microscopic living material such
as DNA.
• These pores or openings are just big enough to fit a single strand of
DNA.
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4. NANOPORESequencing Device
Portable analysis using Flongle and MinION
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Flongle GridION
MinION
High-throughput benchtop
sequencing onGridION
MinIONStarter Packs are available from just $1,000 providing low-cost access to the benefits of long-read,
real-time DNA sequencing.

5. What’s Inside the FLOWCELL
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1. NANOPORE
A protein nanopore is set in an electrically-resistant polymer
membrane.
Types of Nanopores:
A. Biological Nanopore
B. SolidState Nanopore
• Oxford Nanopore's first generation of technology uses pore-forming proteins to create pores in membranes.
• For example, the protein α-hemolysin and similar protein pores are found naturally in cell membranes, where they act as
channels for ions or molecules to be transported in and out of cells.

6. • α-Hemolysin is a heptameric protein pore with an inner diameter of 1 nm, about 100,000 times smaller than
that of a human hair.
• This diameter is the same scale as many single molecules, including DNA.
• The pore is highly stable and has been characterized in great detail .
• Specific adaptations can be designed so that the nanopore is a sensor for a range of specific molecules.
Techniques include:
1. Changing the architecture of the internal structure of the nanopore so that it affects the passage
of an analyte through the pore.
2. The incorporation of a DNA probe to detect an organism with the matching DNA code.
3. The attachment of a molecular motor – for example a processive enzyme – for the analysis of
polymers such as DNA.
4. The attachments of ligands/aptamers to the nanopore, to bind with target proteins outside the
pore.
Biological Nanopores
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7. • A solid-state nanopore is typically a nanometer-sized hole
formed in a synthetic membrane (usually SiNx or SiO2).
• Oxford Nanopore has ongoing R&D and intellectual
property in sold state nanopores.
• Graphene is a robust, single-atom-thick ‘honeycomb’
lattice of carbon with high electrical conductivity.
• By Garaj S et al., Nature 467 (7312), 190–193 (2010) the
Branton andGolovchenko teams used graphene to
separate two chambers containing ionic solutions, and
created a nanopore in the graphene.
SolidState Nanopores
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8. Each microscaffold supports a membrane and embedded nanopore.
The array keeps the multiple nanopores stable during shipping and
usage.
2.Array of microscaffolds :
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3. Sensor chip :
Each microscaffold corresponds to its own electrode that is connected
to a channel in the sensor array chip.Sensor arrays may be
manufactured with any number of channels.

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Working Principle
• Nanopore-based sequencing technology detects the unique electrical signals of
different molecules as they pass through the nanopore with a semiconductor-based
electronic detection system.
• This technology makes for a high throughput, cost effective sequencing solution.
• At the heart of the technology is the biological nanopore, a protein pore embedded in a
membrane, while the brains of the technology lie in the electronics of a semiconductor
integrated circuit and proprietary chemistries.

10. Methodology
Scientific findings 10

11. Methodology
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12. How Does NANOPOREWorks ??
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13. Thank you
KuldepGauliya
206-555-0146
kuldeepgauliya11@gmail.com
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