This presentation provides an overview of next generation sequencing (NGS). It discusses the history of DNA sequencing, from early methods like Maxam-Gilbert sequencing and Sanger sequencing, to newer technologies like 454 sequencing, Illumina sequencing, and Ion Torrent sequencing. These NGS methods allow for massively parallel sequencing of many DNA fragments to generate millions of short reads. The presentation outlines the types of NGS instruments and their applications in fields like molecular biology, medicine, and forensics. It also discusses future challenges in DNA sequencing and decreasing costs to analyze more human genomes.

1. Represented by-
Ajeet kr. Maurya
B.Tech (biotech.)
I.E.T. Bundelkhand
University, Jhansi
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2. PRESENTATIO
N
ON
NEXT
GENERATION
SEQUENCING

3. CONTENT
 Introduction
 History of DNA sequencing
 Types of sequencing
 Next Generation Sequencing(NGS)
 Types of NGS instrument
 Application of NGS
 Future challenges
 Conclusion
 Reference
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4. INTRODUCTION
 DNA Sequencing: Process of determining
the precise order of nucleotide within
a DNA molecule
 It includes any method or technology that
is used to determine the order of four bases-
 Adenine
 Guanine
 Cytosine
 Thymine
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6. TYPES OF SEQUENCING
BASIC METHOD
 Maxam-Gilbert sequencing
 Sanger sequencing
ADVANCED METHOD
 Shotgun sequencing
 Bridge PCR
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7. MAXAM-GILBERT
SEQUENCING
 Base modification
 Removal of modified
base from its sugar
 Breaking the phosphodiester
bond
 Analyzing the fragment using
gel electrophoresis
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9. NEXT GENERATION SEQUENCING
 Tradition (Sanger) sequencing generates a small number
of intermediate length reads(~1000 bp)
 All NGS technologies perform millions of parallel sequencing
reactions to many,typically short,reads per run
 High-throughput sequencing
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Next Generation Sequencing: Why
Now?
• Motivation: HGP and its derivatives, personalized medicine
• Short reads applications: (re-)sequencing, other methods (e.g. gene expression)
• Advancements in technology

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12. TYPE OF NGS METHOD
 454 Sequencing
 ABI’s SOLiD
 Illumina Technology(Solexa)
 Ion Torrent Sequencing
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13. 454 SEQUENCING13

14. ABI’s SOLiD
• Probably the second most widely used
• The workflow is similar to Solexa/Illumina’s
• An intensity difference: SoLiD uses a di-base
sequencing technique in which two nucleotide
are read simultaneously,16 di-base still repeated
by 4 “colors” but the one base shift the redundancy
• As a consequence:
- Sequencing error may propagate
- Read alignment can be speed up
• Error rate around 2-4%
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18. COMPARISON OF NGS METHOD18

19. APPLICATION OF NGS
 Molecular biology
 Evolutionary biology
 Metagenomics
 Medicine
 Forensics
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20. FUTURE CHALLENGES
 Defining variability in many human beings
 DNA sequencing currently under development
include reading the sequencing as a DNA strands
transits through nanopores
 Third generation technologies aim to increase
throughput and decrease the time to result and cost
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21. CONCLUSION
 The sequence based characterization of genome is a relatively
young pursuit in biological science, enhanced model
organism and human genetics
 Ability to gather genome-wide sequence information more rapidly
that can inform a higher level appreciation of functional genome
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22. REFERENCE
Elaine R.Mardis(2007) The impact of next generation sequencing
technology on genetics ,Washington University,U.S.A,133-141
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