DNA sequencing is a method to determine the order of nucleotide bases (adenine, guanine, cytosine, thymine) in a DNA molecule. The Sanger method is more efficient than previous methods as it uses fewer toxic chemicals and radioactive materials. It involves priming DNA with a primer, dividing the primed DNA into four tubes with a DNA polymerase and one of four dideoxynucleotides. These terminate DNA strand elongation randomly, generating fragments of different lengths that can be separated by gel electrophoresis to reveal the DNA sequence. DNA sequencing has applications in forensics, medicine, and agriculture.
Pyrosequencing is a method of DNA sequencing (determining the order of nucleotides in DNA) based on the "sequencing by synthesis" principle, in which the sequencing is performed by detecting the nucleotide incorporated by a DNA polymerase. Pyrosequencing relies on light detection based on a chain reaction when pyrophosphate is released. Hence, the name pyrosequencing.
The chain-termination method developed by Frederick Sanger and coworkers in 1977. This method used fewer toxic chemicals and lower amounts of radioactivity than the Maxam and Gilbert method. Because of its comparative ease, the Sanger method was soon automated and was the method used in the first generation of DNA sequencers.
Pyrosequencing is a method of DNA sequencing (determining the order of nucleotides in DNA) based on the "sequencing by synthesis" principle, in which the sequencing is performed by detecting the nucleotide incorporated by a DNA polymerase. Pyrosequencing relies on light detection based on a chain reaction when pyrophosphate is released. Hence, the name pyrosequencing.
The chain-termination method developed by Frederick Sanger and coworkers in 1977. This method used fewer toxic chemicals and lower amounts of radioactivity than the Maxam and Gilbert method. Because of its comparative ease, the Sanger method was soon automated and was the method used in the first generation of DNA sequencers.
Sanger sequencing is a method of DNA sequencing based on the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication.
Next Generation Sequencing (NGS) Is A Modern And Cost Effective Sequencing Technology Which Enables Scientists To Sequence Nucleic Acids At Much Faster Rate. In This Presentation, You Will Learn About What is NGS, Idea Behind NGS, Methodology And Protocol, Widely Adapted NGS Protocols, Applications And References For Further Study.
DNA Sequencing : Maxam Gilbert and Sanger SequencingVeerendra Nagoria
DNA sequencing is a technique to find out the exact arrangement of Nucleotides to make one strand of DNA. DNA sequencing helps in numerous ways from sequence information to paternity testing, mutation detection etc. Traditionally two approaches were used to solve the problem. First is based of enzymes and Second is based on ddNTPs to sequence the DNA using gel electrophoresis technique.
Probes are used for hybridization purposes. different types of probes can be used on the basis of what we want to hybridize. May be Radioactive or Non-Radioactive.
Sanger sequencing is a method of DNA sequencing based on the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication.
Next Generation Sequencing (NGS) Is A Modern And Cost Effective Sequencing Technology Which Enables Scientists To Sequence Nucleic Acids At Much Faster Rate. In This Presentation, You Will Learn About What is NGS, Idea Behind NGS, Methodology And Protocol, Widely Adapted NGS Protocols, Applications And References For Further Study.
DNA Sequencing : Maxam Gilbert and Sanger SequencingVeerendra Nagoria
DNA sequencing is a technique to find out the exact arrangement of Nucleotides to make one strand of DNA. DNA sequencing helps in numerous ways from sequence information to paternity testing, mutation detection etc. Traditionally two approaches were used to solve the problem. First is based of enzymes and Second is based on ddNTPs to sequence the DNA using gel electrophoresis technique.
Probes are used for hybridization purposes. different types of probes can be used on the basis of what we want to hybridize. May be Radioactive or Non-Radioactive.
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
Gene Sequencing, a tool to analyze the exact order of nucleotide sequence in the DNA -Deoxyribonucleic Acid.
Focuses on Two methods:
a. Maxam-Gilbert (Chemical Degradation) Method
b. Sanger's Method (Dideoxy Chain termination Method)
How to Create Map Views in the Odoo 17 ERPCeline George
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For more information, visit-www.vavaclasses.com
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How to Split Bills in the Odoo 17 POS ModuleCeline George
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Unit 8 - Information and Communication Technology (Paper I).pdfThiyagu K
This slides describes the basic concepts of ICT, basics of Email, Emerging Technology and Digital Initiatives in Education. This presentations aligns with the UGC Paper I syllabus.
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Model Attribute Check Company Auto PropertyCeline George
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2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
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Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
Students, digital devices and success - Andreas Schleicher - 27 May 2024..pptxEduSkills OECD
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2. What Is DNA Sequencing ?
The term DNA Sequencing refers to method for
determining the order of the nucleotide bases
Adenine, Guanine, Cytosine and Thymine in a
molecule of DNA
3. WHY DNA SEQUENCING IS
IMPORTANT ?
• The precise usage of codons, information
regarding mutations and polymorphisms and
the identification of gene regulatory control
sequence can only be elucidated by analyzing
DNA sequence
4.
5. Disadvantages Of Maxam & Gilbert
Chemical Degradation Method
• Requires lots of purified DNA and many
intermediate purification step
• It requires extensive use of hazardous
chemicals
• It is difficult to scale up and cannot be used to
analyze more than 500bp
6.
7. SANGERS METHOD OF DNA
SEQUENCING
• The chain terminator method is more efficient
and uses fewer toxic chemicals and lower
amount of radioactivity than the method of
Maxam and Gilbert.
• The key principle of the Sangers method was
use of dideoxynucleotides triphosphate
(ddNTPs) as DNA chain terminator
8.
9.
10.
11. Chain termination method of DNA
sequencing
• It involves following components:
1. Primer
2. DNA template
3. DNA polymerase
4.dNTPs(A,T,G,C)
5. ddNTPs
• It involves following 4 Steps:
1. Denaturation
2. Primer attachment and extension of bases
3. Termination
4. Poly acrylamide gel electrophoresis
12. • The Methods (Procedure ):-
1. Before the DNA can be sequenced, it has to be
denatured into single strands using heat.
2. Next a primer is annealed to one of the template
strands. This primer is specifically constructed so that
its 3' end is located next to the DNA sequence of
interest. Either this primer or one of the nucleotides
should be radioactively or fluorescently labeled so
that the final product can be detected on a gel. Once
the primer is attached to the DNA, the solution is
divided into four tubes labeled "G", "A", "T" and "C".
13. • ‘’G’’ tubes : all four dNTP’s, ddGTP and DNA
polymerase
• ‘’A’’ tubes : all four dNTP’s, ddATP and DNA
polymerase
• ‘’T’’ tubes : all four dNTP’s, ddTTP and DNA
polymerase
• ‘’C’’ tubes : all four dNTP’s, ddCTP and DNA
polymerase
14.
15. • Mixture first heated so that DNA strands separate
(96⁰C)
• Then temperature get lower so that short length DNA
sequence – a primer can bind to the template
DNA(50⁰C)
• Temperature raised to (60⁰C- 65⁰C)to enable the DNA
Polymerase enzyme to bind to the short section of
double stranded DNA.
• DNA polymerase to synthesizes new DNA. DNA
Polymerase will continue adding Nucleotides to chain
until it happens to add a dideoxy nucleotide instead of
normal one
16.
17.
18. Uses of Sangers sequence
• Region up to about 900 basepairs in length
sequenced using this method
• In Human Genome project sanger sequencing
was used to determine the sequence of
relatively small fragments of human DNA
• Sanger sequencing is still in wide use for the
sequencing of individual pieces of DNA, such
as fragments used in DNA cloning
19. • Advantage of Basic method
• 1:-Improvement diagnosis of disease.
• 2:- Identifying suspects .
• Disadvantage :
• 1:-Whole genome can not be sequenced at
once .
• 2:- Very slow and time consuming
20.
21. • Applications of DNA
Sequencing
• Forensics: to help identify individuals because
each individual has a different genetic sequence
• Medicine: can be used to help detect the genes
which are linked to various genetic disorders such
as muscular dystrophy.
• Agriculture: The mapping and sequencing of a
genome of microorganisms has helped to make
them useful for crops and food plants.
