WHAT IS DNA SEQUENCER ?
DNA SEQUENCING.
HISTORY
MANUFACTURERES OF DNA SEQUENCER.
OTHER APPLICATIONS
CONCLUSION
REFERENCES
The order of the DNA bases is reported as a text string, called a read. Some DNA sequencers can be also considered optical instruments as they analyze light signals originating from fluorochromes attached to nucleotides.
1. DNA SEQUENCER
By
KAUSHAL KUMAR SAHU
Assistant Professor (Ad Hoc)
Department of Biotechnology
Govt. Digvijay Autonomous P. G. College
Raj-Nandgaon ( C. G. )
2. SYNOPSIS
WHAT IS DNA SEQUENCER ?
DNA SEQUENCING.
HISTORY
MANUFACTURERES OF DNA
SEQUENCER.
OTHER APPLICATIONS
CONCLUSION
REFERENCES
3. What is DNA sequencer ?
A DNA sequencer is a scientific instrument used
to automate the DNA sequencing process.
Given a sample of DNA, a DNA sequencer is used
to determine the order of the four bases: adenine,
guanine, cytosine, and thymine.
The order of the DNA bases is reported as a text
string, called a read. Some DNA sequencers can
be also considered optical instruments as they
analyze light signals originating from
4. DNA sequencing
Determination of nucleotide sequence in a
DNA molecule is known as DNA sequencing.
This is the basic and fundamental
requirement in biotechnology.
DNA sequencing is important to understand
the function of genes, and basis of inharited
disorders.
Also, DNA cloning and gene manipulation
invariably require knowledge of accurate
5. History about DNA sequencing
The first DNA sequencing methods
were developed by Gilbert (1973) and
Sanger (1975).
Gilbert introduced a sequencing method
based on chemical modification of DNA
followed by cleavage at specific bases
whereas Sanger’s technique is based
on dideoxynucleotide chain termination.
The Sanger method became popular
6. History about DNA sequencer
The first automated DNA sequencer was
introduced by Applied Biosystems in 1987.
It used the Sanger sequencing method, a
technology which formed the basis of the “first
generation” of DNA sequencers and enabled
the completion of the human genome project
in 2001.
The human genome project catalysed the
development of cheaper, high throughput and
more accurate platforms known as Next
Generation Sequencers (NGS).
These include the 454, SoLiD (Supported
7. Next generation sequencing machines have
increased the rate of DNA sequence
substantially compared with previous Sanger
methods.
DNA samples can be prepared automatically
in as little as 90 mins, while a human genome
can be sequenced at 15 times coverage in a
matter of days.
More recent, third-generation DNA
sequencers such as SMRT (Single-molecule
real-time) and Oxford Nanopore measure the
addition of nucleotides to a single DNA
Continue….
8. Manufacturers of DNA sequencers
DNA sequencers have been developed,
manufactured, and sold by the following
companies, among others.
ROCHE
ILLUMINA
LIFE TECHNOLOGIES
BACKMAN COULTER
PACIFIC BIOSCIENCE
9. Roche
The 454 DNA sequencer was the first next-
generation sequencer to become
commercially successful.
It was developed by 454 Life Sciences and
purchased by Roche in 2007.
454 utilizes the detection of pyrophosphate
released by the DNA polymerase reaction
when adding a nucleotide to the template
strain.
10. Continue.......
Roche currently manufactures two systems based
on their pyrosequencing technology: the GS FLX+
and the GS Junior System.
The GS FLX+ System promises read lengths of
approximately 1000 base pairs while the GS Junior
System promises 400 base pair reads.
A predecessor to GS FLX+, the 454 GS FLX
Titanium system was released in 2008 , achieving
an output of 0.7G of data per run, with 99.9%
accuracy after quality filter, and a read length of up
to 700bp.
In 2009, Roche launched the GS Junior, a bench
top version of the 454 sequencer with read length
up to 400bp, and simplified library preparation and
12. ILLUMINA
Illumina produces a number of next-generation
sequencing machines using technology acquired
from Manteia Predictive Medicine and developed
by Solexa.
Illumina makes a number of next generation
sequencing machines using this technology
including the HiSeq, Genome Analyzer IIx, MiSeq
and the HiScanSQ, which can also process
microarrays.
13. sequencers was first released by Solexa in
2006 as the Genome Analyzer.
Illumina purchased Solexa in 2007.
The Genome Analyzer uses a sequencing by
synthesis method. The first model produced
1G per run.
During the year 2009 the output was
increased from 20G per run in August to 50G
per run in December.
In 2010 Illumina released the HiSeq 2000
with an output of 200 and then 600G per run
which would take 8 days.
At its release the HiSeq 2000 provided one of
the cheapest sequencing platforms at $0.02
per million bases as costed by the Beijing
Genomics Institute.
14. Life Technologies
Life Technologies produces DNA sequencers
under the Applied Biosystems and Ion Torrent
brands.
Applied Biosystems makes the SOLiD next-
generation sequencing platform, and Sanger-
based DNA sequencers such as the 3500
Genetic Analyzer.
Under the Ion Torrent brand, Applied
Biosystems produces two next-generation
sequencers: the Ion PGM System and the Ion
Proton System.
15. Beckman Coulter
Beckman Coulter (now Danaher) has
previously manufactured chain termination
and capillary electrophoresis-based DNA
sequencers under the model name CEQ,
including the CEQ 8000.
The company now produces the GeXP
Genetic Analysis System, which uses dye
terminator cycle sequencing.
This method uses a thermocycler in much
the same way as PCR to denature, anneal,
and extend DNA fragments, amplifying the
sequenced fragments.
16. Pacific Biosciences
Pacific Biosciences produces a
sequencing system named the PacBio RS
using a single molecule real time
sequencing, or SMRT, method.
This system can produce read lengths of
multiple thousands of base pairs, though
with a high rate of errors.
These errors may be alleviated by use of
optimized assembly strategies.Scientists
have reported 99.999% accuracy with these
strategies.
17. Other applications
DNA sequencers exert a precise control over
the temperature of individual reactions (wells) in
reaction plates.
This fact, combined with a fluorescence
readout capability and their ubiquitous presence
in biology laboratories, suggests that DNA
sequencers may be used to study temperature
dependent events in cells.
For example, this approach was recently used
in the study of the temperature-activated ion
channel TRPV1.