Next generation DNA sequencing refers to modern massively parallel DNA sequencing technologies that can generate millions of sequences simultaneously. It involves sequencing small DNA fragments in parallel and then bioinformatics to assemble the sequences. Major NGS platforms include pyrosequencing (Roche 454), Illumina sequencing, and Ion Torrent semiconductor sequencing. Each has advantages like high throughput and lower costs but also limitations such as shorter read lengths or errors in homopolymers. The first human genome cost over $2 billion but can now be sequenced for around $1000.
In a detail description of the two major blotting techniques. Right from its history to the result interpretation put forth in a concise way. Helps understand these protocols with ease.
In a detail description of the two major blotting techniques. Right from its history to the result interpretation put forth in a concise way. Helps understand these protocols with ease.
The next generation sequencing platform of roche 454creativebiogene1
454 is totally different from Solexa and Hiseq of Illumina. The disadvantage of 454 is that it is unable to accurately measure the homopolymer length. For this unavoidable reason, 454 technology will introduce insertion and deletion sequencing errors to the results.
This presentation is explains about the genome sequencing, its traditional method and modern method. This basically focus on Next Generation Sequencing and its types.
It contains information about- DNA Sequencing; History and Era sequencing; Next Generation Sequencing- Introduction, Workflow, Illumina/Solexa sequencing, Roche/454 sequencing, Ion Torrent sequencing, ABI-SOLiD sequencing; Comparison between NGS & Sangers and NGS Platforms; Advantages and Applications of NGS; Future Applications of NGS.
Sequencing is one of the major technological advancement that has taken shape in the last two or three decade. Starting from Sanger and Maxam-Gilbert sequencing methods to the latest high-throughput methods, sequencing technologies has changed the the landscape of biological sciences.
This slide takes a look a the major sequencing methods over time.
Note: Several images included here have been sourced from GOOGLE IMAGES. The content has been extracted from several SCIENTIFIC PAPERS and WEBSITES.
PLEASE DO CONTACT THE AUTHOR DIRECTLY IF ANY COPYRIGHT ISSUE ARISES.
Detailed explanation about gene sequencing methods
Sequencing the gene is an important step toward understanding the gene.
A gene sequence contains some clues about where genes are.
Gene sequencing give us understanding how the genome as a whole works-how genes work together to direct the growth, development and maintenance of an entire organism.
It help scientists to study the part of genome outside the genes-regulatory regions
Deciphering DNA sequences is essential for virtually all branches of biological research. With the
advent of capillary electrophoresis (CE)-based Sanger sequencing, scientists gained the ability to
elucidate genetic information from any given biological system. This technology has become widely
adopted in laboratories around the world, yet has always been hampered by inherent limitations in
throughput, scalability, speed, and resolution that often preclude scientists from obtaining the essential
information they need for their course of study. To overcome these barriers, an entirely new technology
was required—Next-Generation Sequencing (NGS), a fundamentally different approach to sequencing
that triggered numerous ground-breaking discoveries and ignited a revolution in genomic science.
The next generation sequencing platform of roche 454creativebiogene1
454 is totally different from Solexa and Hiseq of Illumina. The disadvantage of 454 is that it is unable to accurately measure the homopolymer length. For this unavoidable reason, 454 technology will introduce insertion and deletion sequencing errors to the results.
This presentation is explains about the genome sequencing, its traditional method and modern method. This basically focus on Next Generation Sequencing and its types.
It contains information about- DNA Sequencing; History and Era sequencing; Next Generation Sequencing- Introduction, Workflow, Illumina/Solexa sequencing, Roche/454 sequencing, Ion Torrent sequencing, ABI-SOLiD sequencing; Comparison between NGS & Sangers and NGS Platforms; Advantages and Applications of NGS; Future Applications of NGS.
Sequencing is one of the major technological advancement that has taken shape in the last two or three decade. Starting from Sanger and Maxam-Gilbert sequencing methods to the latest high-throughput methods, sequencing technologies has changed the the landscape of biological sciences.
This slide takes a look a the major sequencing methods over time.
Note: Several images included here have been sourced from GOOGLE IMAGES. The content has been extracted from several SCIENTIFIC PAPERS and WEBSITES.
PLEASE DO CONTACT THE AUTHOR DIRECTLY IF ANY COPYRIGHT ISSUE ARISES.
Detailed explanation about gene sequencing methods
Sequencing the gene is an important step toward understanding the gene.
A gene sequence contains some clues about where genes are.
Gene sequencing give us understanding how the genome as a whole works-how genes work together to direct the growth, development and maintenance of an entire organism.
It help scientists to study the part of genome outside the genes-regulatory regions
Deciphering DNA sequences is essential for virtually all branches of biological research. With the
advent of capillary electrophoresis (CE)-based Sanger sequencing, scientists gained the ability to
elucidate genetic information from any given biological system. This technology has become widely
adopted in laboratories around the world, yet has always been hampered by inherent limitations in
throughput, scalability, speed, and resolution that often preclude scientists from obtaining the essential
information they need for their course of study. To overcome these barriers, an entirely new technology
was required—Next-Generation Sequencing (NGS), a fundamentally different approach to sequencing
that triggered numerous ground-breaking discoveries and ignited a revolution in genomic science.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
2. Hello!
I am Mousumee Mahapatra
Department of Botany
Guru Ghasidas Vishwavidyalaya (CU)
Bilaspur, Chhattisgarh
3. What Is Next Generation Sequencing?
⊹ In 2005 , new era of DNA sequencing have emerged , when a fully
automated massively parallel pyrosequencing machine was
developed . This led to foundation of Next generation Sequencing.
⊹ It is a choice for large scale genomic and transcription
sequencing because of high outputs (Gigabyte per instrument)
range.
⊹ It involves the sequencing of small DNA fragments in parallel
and then use of bioinformatics analysis to arrange the sequence
of those fragments together either by mapping the individual
reads to reference genome.
⊹ Thus it is also called massive parallel sequencing or deep
sequencing.
4. “
⊹ Work flow of NGS
Video source- https.//info.abmgood.com/next-generation-
sequencing
5. Types of next generation sequencing
Pyrosequencing
Non-electrophoretic,
bioluminescence method
that measures the release
of inorganic pyrophosphate
by propertionally
converting it into visible
light using a series of
enzymatic option.
Illumina sequencing
Illumina sequencing
technology leverages clonal
array formation and
proprietary reversible
terminator technology for
rapid and accurate
large-scale sequencing.
Ion-semiconductor
sequencing
Ion semiconductor sequencing is a
method of DNA sequencing based on
the detection of hydrogen ions that
are released during the
polymerization of DNA. This is a
method of "sequencing by synthesis",
during which a complementary strand
is built based on the sequence of a
template strand.
5
7. Why
Pyrosequencing?
7
Pyros (Greek for “fire,” because light is produced) because genome
sequencing is done utilizing light-emitting enzyme- Luciferase.
Luciferase emits lights in presence of ATP present in several
organisms such as American firefly and poisonous Jack-o-lantern
mushroom.
8. WORK FLOW OF PYROSEQUENCING
8
Video source- https.//info.abmgood.com/next-generation-
sequencing
9. How it works?
⊹ In DNA synthesis, a dNTP is attached to the 3’end
of the growing DNA strand. The two phosphates
on the end are released as pyrophosphate (PPi).
⊹ ATP sulfurylase uses PPi and adenosine 5’-
phosphosulfate to make ATP.
⊹ Luciferase is the enzyme that causes fireflies to
glow. It uses luciferin and ATP as substrates
converting luciferin to oxyluciferin and releasing
visible light.
– The amount of light released is proportional to
the number of nucleotides added to the new DNA
strand.
⊹ After the reaction has completed, apyrase is added
to destroy any leftover dNTPs. 9
10. Procedure involved in pyrosequencing
⊹ DNA is fragmented: To start, the DNA is sheared into 300-800 bp
fragments, and the ends are “polished” by removing any unpaired
bases at the ends.
⊹ The DNA strand’s ends are made blunt with appropriate enzymes
⊹ “A” and “B” adapters are ligated to the blunt ends using DNA
ligase
⊹ The strands are denatured using sodium hydroxide to release the
ssDNA template library (sstDNA).
The adapters
⊹ The A and B adapters are used as priming sites for both
amplification and sequencing since their composition is known.
⊹ The B adapter contains a 5’ biotin tag used for mobilization.
⊹ The beads are magnetized and attract the biotin in the B adaptors.
1. Preparation of Sample DNA
http://www.pyrosequencing.com/DynPage.aspx?id=7454
11. ⊹ Using water-in-oil emulsion, each ssDNA in the library is hybridized onto
a primer coated bead.
⊹ By limiting dilution, an environment is created that allows each emulsion
bead to have only one ssDNA.
⊹ Each bead is then captured in a its own emulsion micro-reactor,
containing in it all the ingredients needed for a PCR reaction.
⊹ PCR takes place in each of these beads individually, but all in parallel.
⊹ This activity as a whole is emPCR.
One adapter contains biotin, which binds to a streptavidin-coated bead.
The ratio of beads to DNA molecules is controlled so that most beads get
only a single DNA attached to them.
Oil is added to the beads and an emulsion is created. PCR is then
performed, with each aqueous droplet forming its own microreactor.
Each bead ends up coated with about a million identical copies of the
original DNA
11
2. Cloning of DNA
http://www.pyrosequencing.com/DynPage.aspx?id=7454
12. ⊹ Utilizing the A adapter, a primer is added to the ssDNA.
⊹ The beads are now loaded into individual wells created from finely packed and cut fiber-optics (PicoTiterPlate
device).
⊹ The size of the wells do not allow more than one ssDNA bead to be loaded into a well.
⊹ Enzyme beads and packing beads are added. Enzyme beads containing sulfurase and luciferase, and packing
beads used only to keep the DNA beads in place.
⊹ Above the wells is a flow channel, passing nucleotides and apyrase in a timed schedule.
⊹ Comparing the peak light emission of incorporation of C or T at a CpG side with in the amplicon gives a precis3
measure of amount of methylation at the position within the sample.
12
3. Sequencing
https://www.researchgate.net/figure/The-principle-of-Pyrosequencing-a-method-of-sequencing-by-synthesis-from-22-
The_fig2_267972290?hcb=1
13. advantages
13
1 2 3
3
2
1
Larger sequence can be read easily. Approx.
48,000 sequencing can be done per day It is a much accurate
Easily automated and
there is no need of gel
electrophoresis.
High reagent costs. Thus
not economic friendly.
There is high error rate
over strings of 6+
homopolymer.
The work required is
considerable, and special
software is required for the
instrument
and for the analysis.
Disadvantages
15. Why Illumina
sequencing
15
• It is the most successful sequencing system with a claimed >70%
dominance of the market.
• The Illumina sequencer is different from the Roche 454
(Pyrosequencing) sequencer in that it adopted the technology of
sequencing by synthesis using removable fluorescently labeled
chain-terminating nucleotides that are able to produce a larger
output at lower reagent cost
16. Work frame of illumina sequencing
16
Video source- https.//www.illumine.com/next-generation-sequencing
17. Procedure involved in illumine sequencing.
⊹ Fragment DNA of interest into smaller strands that are able to
be sequenced.
• Sonication
• Nebulization
• Enzyme digestion
⊹ Ligate Adapters.
⊹ Denature dsDNA into ssDNA by heating to 95° C.
1. Preparation of Genomic DNA sample
18. ⊹ ssDNA is then bound to inside surface of flow
cell channels
⊹ Dense lawn of primer on the surface of the flow
cell
2. Attach DNA to surface
⊹ Unlabeled nucleotides and polymerase enzyme
are added to initiate the solid phase bridge
amplification
3. Bridge amplification
19. ⊹ In this step it demonstrates the work done by the
sequencing reagents
• Primers
• Nucleotides
• Polymerase enzymes
• Buffer
19
4. Fragments become double stranded
20. ⊹ The original strand is then washed away,
leaving only the strands that had been
synthesized to the oligos attached to the
flow cell
20
5. Denature the double stranded molecules.
Attached
Attached
6. Complete amplification
Cluster
⊹ Cycle of new strand synthesis and Denaturation to make
multiple copies of the same sequence (amplification)
• Fragments Become Double Stranded
• Denature the Double Strand Molecules
21. ⊹ The P5 region is cleaved
⊹ Add sequencing reagents
• Primers
• Polymerase
• Fluorescently labelled nucleotides
• Buffer
⊹ First base incorporated
7. Determine first base
22. ⊹ Remove unincorporated bases
• Detect Signal
• Deblock and remove the
⊹ Fluorescent signal new cycle
22
8. Image first base
⊹ Add sequencing reagents
• Primers
• Polymerase
• Fluorescently labeled nucleotides
• Buffer
⊹ Second base incorporated
9. Determine second base
23. 23
⊹ The identity of each base of a cluster is
read off from sequential images
11. Sequence Reads Over Multiple Chemistry Cycles
10. Image second chemistry cycle
⊹ Remove unincorporated bases
• Detect Signal
• Deblock and remove the
⊹ Fluorescent signal new cycle
24. 24
12. Align data
⊹ After the sequencing is finished they are aligned.
⊹ Each was once one larger sequence that had been
fragmented.
⊹ Needs to be realigned to find the original sequence
of the larger sequence.
25. Application of illumina sequencing
Sequence based
transcriptome
analysis
SNPs and SVs
discovery and small
RNA discovery
analysis.
Cytogenetic analysis
and ChIP sequencing
2 4
DNA sequencing gene
regulation analysis
1 3
26. Merits and demerits of illumine sequencing
Merits :
⊹ Good for sequencing
of repetitive and
homopolymer
sequences .
⊹ Long read length
⊹ Preferable for Whole
genome sequencing
Demerits :
⊹ Incomplete removal
of fluorescent
molecules create
high background
noise .
26
28. Why ion-semiconductor
sequencing?
28
• It is based on release of proton (H+) after nucleotide
incorporation .
• Sequencing chip contains many small wells . Ideally ,
each well contains single bead with clonally
amplified DNA (cluster) ( i.e. – amplified in ePCR ) .
• Each cluster located directly above a semiconductor
transistor which is capable to determine change in
pH of solution.
29. Work frame of ion-semiconductor sequencing
29
Video source- https.//info.abmgood.com/next-generation-sequencing
30. advantages
Less reagent
cost .
30
No need of
chemically
modified
nucleotides .
No need of
nucleotides .
disadvantages
Short reads High error rate
over
homopolymer
31. Cost of sequencing and other specifications of different NGS
platforms :
31
* RP = reagent price + sequencing price
NA = Not Available
Source- https://www.longdom.org/open-access/generations-of-sequencing-technologies-from-first-to-
next-generation-0974-8369-1000395.pdf
32. COST OF DNA SEQUENCING :
32
Source- https://www.genome.gov/about-genomics/fact-sheets/Sequencing-Human-
Genome-cost
33. How much was the first human genome sequence .
⊹ The total cost of Human genome project (HGP) [ from October 1990 to April 2003 ]
was estimated as ~ $ 2.7 billion .
⊹ The estimated cost of first draft sequence ( i.e.- ~ 90% coverage of genome at
99.9% accuracy ) was ~ $ 300 million worldwide . ( April 1999 - June 2000)
⊹ The finished sequence ( > 95% coverage of genome at 99.99% accuracy ) was
submitted on 2003 ; of which the estimated cost was ~$150 million worldwide .
⊹ NHGRI estimated that the hypothetical 2003 cost to generate a second reference
genome sequence using then-available approaches and technologies was in the
neighborhood of $ 50 million .
⊹ At present , HiSeq X Ten System , released in 2014 , can sequence over 45 human
genomes in a single day for approximately $ 1000 each .
33
