Next-Gen

Ion Torrent Sequencing

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by gbnita
Last updated 1 year ago

Discipline:
Science
Subject:
Experiments

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Ion Semiconductor Sequencing Technology

Binita Giri and Sajjad Abedian

Next Generation Sequencing Technologies have seen an exponential growth in recent years. These sequencing technologies run for a variety of applications such as whole genome sequencing, exome sequencing, targeted sequencing, transcriptome sequencing, chip-seq, RNA seq, etc. Currently, there are number of technologies which perform these analysis. One of the latest method to the NGS marketplace is 'Ion Torrent Semiconductor' which was introduced in 2010 by the company called 'Life Technologies'.Ion Torrent combines semiconductor sequencing technology and biochemistry to convert chemical information (A, C, G, T) into digital data (0, 1). Unlike many other technologies, this method doesn't use any modified nucleotides or optical technology. Instead, Ion torrent uses complementary metal-oxide semiconductors (CMOS) as used in digital cameras, laptops and smart phones. It uses an electrochemical detection system such as ion-sensitive field-effect transistors (ISEFT) which detects hydrogen ion as they released during DNA polymerization. The release of hydrogen ion causes a pH shift which is detected by CMOS sensor.Ion torrent sequencing is revolutionary among other NGS technology as it is simple, fast, scalable and cost effective.

1. Sample preparation for this method is similar to other technologies like 454 which uses emulsion PCR. Genomic DNA are fragmented, ligated to adapters and these adapters are clonally amplified into beads. Ion torrent sequences large number of these amplified beads as templates on a solid surface.2. In this method, semiconductor chip is the machine. Each chip has around 1.2 million sensors and are composed of well that contains dNTP and an acrylamide bead with a DNA template.3. If the base in DNA is complementary to the nucleotide, a nucleotide will be incorporated and hydrogen ion will be released4. The CMOS layer lies beneath the well which lies over a sensor plate transmits the pH changes to the semiconductor chip. This change is recorded and the base is called. This process occurs simultaneously in all the wells.

Sequencing Chemistry

Work Flow

1. When nucleotide is incorporated by the polymerase in the DNA molecule, a proton is released and the pH of the well changes.2. If there is no incorporation, there is no pH changes.3. Repeated base pairs incorporation results a pH chnage of 2X greater magnitude.

NGS Applications1. Targeted DNA sequencing 2. Targeted RNA sequencing3. Exome sequencing 4. de novo sequencing 5. Transcriptome sequencing 6. Microbial sequencing7. Viral typing 8. Bacterial typing9. Chip sequencing 10. Aneuploidy and CNV analysis

Ion Torrent Products1. Ion S5 Systems2. Ion AmpliSeq Technology3. Ion Proton System4. Ion PGM System5. Ion Chef System

References 1. https://www.youtube.com/watch?v=WYBzbxIfuKs 2. https://www.youtube.com/watch? v=JHzkYDyMzOg3. https://www.youtube.com/watch?v=xDL5VTITD2Y4. "Next Generation Sequencing For Dummies". N.p., 2016. Web. 11 Feb. 2016.5. "An Introduction To Next Generation Sequencing". N.p., 2016. Web. 11 Feb. 2016.6. https://www.thermofisher.com/us/en/home/brands/ion-torrent.html

Why Ion Torrent Sequencing?


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