Successfully reported this slideshow.
Your SlideShare is downloading. ×

NANOPORE SEQUENCING

Ad
Ad
Ad
Ad
Ad
Ad
Ad
Ad
Ad
Ad
Ad
Upcoming SlideShare
Next Generation Sequencing
Next Generation Sequencing
Loading in …3
×

Check these out next

1 of 13 Ad
Advertisement

More Related Content

Recently uploaded (20)

Advertisement

NANOPORE SEQUENCING

  1. 1. N A N O P O R E S E Q U E N C I N G Kuldeep Gauliya
  2. 2. Why Nanopore ?? • Flexibility and versatility • Single molecule sequencing • Both short and long read lengths • Real-time data generation and analysis • Cost-effectiveness 2
  3. 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. 3
  4. 4. NANOPORESequencing Device Portable analysis using Flongle and MinION 4 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. 5. What’s Inside the FLOWCELL 5 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. 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 6
  7. 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 7
  8. 8. Each microscaffold supports a membrane and embedded nanopore. The array keeps the multiple nanopores stable during shipping and usage. 2.Array of microscaffolds : 8 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.
  9. 9. 9 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. 10. Methodology Scientific findings 10
  11. 11. Methodology 11
  12. 12. How Does NANOPOREWorks ?? 12
  13. 13. Thank you KuldepGauliya 206-555-0146 kuldeepgauliya11@gmail.com 13

×