Ion Torrent sequencing is a next generation sequencing technique that detects hydrogen ions released during DNA polymerization using a semiconductor chip, allowing for DNA sequencing without the need for optical detection. The technique involves fragmenting DNA, attaching fragments to beads, amplifying fragments using emulsion PCR, loading beads into wells on an ion semiconductor chip, and flowing nucleotides to detect pH changes from nucleotide incorporation. Key advantages are faster run times, lower costs, and scalability compared to other NGS methods.

1. ION TORRENT SEQUENCING
Submitted By,
ADITHYA P BALAKRISHNAN
04-AGRMA-01724-2018
M.Sc. (Agri.) first year
Dept. of PBG
C.P.C.A
S.D.A.U
ASSIGNMENT PRESENTSTION ON
COURSE NO: MBB 501
COURSE TITLE: PRINCIPLES OF BIOTECHNOLOGY
Submitted To,
Dr. K. P. PACHCHIGAR
Assistant Professor
College Of Basic Science &
Humanities
S.D.A.U

2. ION TORRENT SEQUENCING IS A NEXT
GENERATION SEQUENCING TECHNIQUE.
First generation sequencing : Sanger sequencing
(capillary electrophoresis based)
NGS : Includes several high-throughput
approaches to sequence DNA. (Massively Parallel
Sequencing)

3. How NGS works?
Source of genome : Genomic DNA/ RT-RNA/ cDNA
Three general steps:
1. Library preparation
2. Amplification
3. Sequencing

4. Three general steps of NGS
KHALID,R. and SABAHUDDIN,A.(2016).Principles,analysis,application and challenges of next
Generation sequencing : a review.(2016).

5. Ion torrent technology development
history
 The technology was licensed from
DNA Electronics Ltd, developed by
Ion Torrent Systems Inc. and released
in February 2010.
 Roche’s 454 Life Sciences is
partnering with DNA Electronics on
the development of a long-read,
high-density semiconductor platform
using this technology.
https://upload.wikimedia.org/wikipedia/commons/thumb/2/20/Life_Technologies_-
_Ion_Proton_%28TM%29.jpg/1280px-Life_Technologies_-_Ion_Proton_%28TM%29.jpg

6. Key historical developments
 Introduction of viable ion-sensitive transistor sensor device by
Bergveld in 1970 (pHFET-sensing pH).
 In 1980s use of semiconductor devices into the specific CMOS
process.
 In 2006 Rothberg invented specific CMOS chip massively parallel
sequencing device by combination of 4o years of pHEFT sensor
development & 454 massively parallel sequencing .

7. BARRY, M. and JONATHAN,M.R.(2012).Review: Progress in ion torrent semiconductor chip based sequencing. Electrophoresis 2012.33,3397-3417.

8. BASIC PRINCIPLE OF ION TORRENT
SEQUENCING
Uses the SEQUENCING BY SYNTHESIS approach
(SBS) .
In this approach a new DNA strand complementary to the
targeted strand is synthesized one base at a time.
A semiconductor chip detects H⁺ ions produced during
DNA polymerization.

9. Sequencing Chemistry Of
Ion Torrent
 The incorporation of
deoxyribonucleotide
triphosphate into a growing
DNA strand causes the
release of hydrogen ion /
proton (H⁺) .
https://upload.wikimedia.org/wikipedia/commons/thumb/1/10/DNTP_n
ucleotide_incorporation_reaction.svg/1280px-
DNTP_nucleotide_incorporation_reaction.svg.png

10. Overview of semiconductor pH sensor
technology
BARRY, M. and JONATHAN,M.R.(2012).Review: Progress in ion torrent semiconductor chip based sequencing. Electrophoresis 2012.33,3397-3417.

11. How the process works?
STEPS:
• DNA fragmentation
• Addition of Adaptors/Index/Barcode/Anchor
• Attachment to beads
• Emulsion PCR for amplification
• Tiny wells of chip (heart of ion torrent) loaded with these beads
• Nucleotides flow sequentially over ion semiconductor chip
• pH change recorded

12. Ion torrent sequencing workflow
https://www.google.com/imgres?imgurl=https%3A%2F%2Fwww.researchgate.net%2Fpublication%2F303500733%2Ffigure%2Ffig1
%2FAS%3A668941772926988%401536499555645%2FIon-Torrent-PGM-sequencing-
workflow.ppm&imgrefurl=https%3A%2F%2Fwww.researchgate.net%2Ffigure%2FIon-Torrent-PGM-sequencing-
workflow_fig1_303500733&docid=Wigr3vKmrWAvMM&tbnid=OpGj2KwVWWVtOM%3A&vet=10ahUKEwjN7_DsvsThAhWVAYgKH
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7_DsvsThAhWVAYgKHcvLDDoQMwhZKBIwEg&iact=mrc&uact=8

13. Overview of sequencing using a
semiconductor pH sensor device
https://www.google.com/imgres?imgurl=x-raw-
image%3A%2F%2F%2Fe4a7086d25939fc2d978782c912cf3d0456b8e620afa61a20aef8ea36e86324a&imgrefurl=https%3A%2F%2Fw
ww.viprbrc.org%2FbrcDocs%2Ftutorials%2FModule1a_NGS_WetLab.pdf&docid=4BbmElQhfMAv5M&tbnid=wp-TJW0Ip-
SaVM%3A&vet=10ahUKEwjN7_DsvsThAhWVAYgKHcvLDDoQMwh1KCEwIQ..i&w=882&h=533&bih=738&biw=1506&q=ion%20tor
rent%20sequencing%20workflow&ved=0ahUKEwjN7_DsvsThAhWVAYgKHcvLDDoQMwh1KCEwIQ&iact=mrc&uact=8

14. Features of Ion-Torrent
No use of optical signals to exploit the addition of dNTPs
Run time : 3 hrs
Clusteral amplification : emulsion PCR
Read length : 100 to 300 bp
Throughput determined by chip size : 10Mb -5 Gb
Cost :$0.63 per Mb with 318 chip

15. Advantages of Ion-Torrent sequencing
Less time requirement
Low cost instrument upgraded through disposable chips
Scalable (10-1000 Mb of data per run)
Medium to low cost per base

16. REFERENCES :
BARRY, M. and JONATHAN,M.R.(2012).Review: Progress
in ion torrent semiconductor chip based sequencing.
Electrophoresis 2012.33,3397-3417.
KHALID, R. and SABAHUDDIN,
A.(2016).Principles,analysis,application and challenges of
next-generation sequencing : a review.(2016).