dna sequencing methods

Dna sequencing

Upload: steffie-dmello

Post on 16-Apr-2017

115 views

Category:

Education

0 download

Report

Download

Embed Size (px):

TRANSCRIPT

Dna sequencing

What is DNA sequencing ? • Determining order of nucleotides

• DNA first discovered by Friedrich Miescher in 1869

• Oswald Avery, Colin Macleod and Maclyn McCarty demonstrated that purified DNA could change one strain of bacteria into another

• In 1953 James Watson and Francis Crick put forward double helix model of DNA

• Foundation of sequencing protein was first laid by Sanger

HISTORY

• The first method for determining DNA sequence involved a location specific primer extension strategy, established by Ray Wu in 1970

• DNA polymerase catalysis and specific nucleotide labelling used

• Between 1970 and 1973 R. Padmahabhan and colleagues that this method can be used to identify any DNA sequence

• Sanger adopted primer extension strategy to develop rapid DNA sequences

• Later Maxam and Gilbert also developed sequencing method

• Why is DNA sequencing important ?

Molecular medicines Bioarchaeology, Anthropology,

Evolution and Human migration

DNA Forensics Detect bacteria and other organisms

that pollute air Agriculture, livestock breeding and

bioprocessing

Maxam-Gilbert method

Sangers dideoxynucleotide method

Bacteriophage M13

Dna

sequ

enci

MAXAM GILBERT METHOD

• Allan Maxim and Walter Gilbert developed a method for sequencing single stranded dna

• Purines dimethyl sulphate • Pyrimidines hydrazine • Breaking of glyosidic bonds between

ribose sugar and base

DimethylSulphate

Piperdine

Guanine

Hydrazine

Piperdine

Thymine + Cytosin

Guanine +

Adenine

Dimethyl sulphate

Piperdine Formic

acid

Cytosine

Hydrazine

Piperdine Nacl

• Discovered by Scientist Sanger • Also know as dideoxy method or

chain termination method • 2 main things to be done :1. Denature the DNA 2. Labelling

SANGER SEQUENCING

DIDEOXYNUCLEOTIDE

• In DNA nucleotides are bond to each other by phosphodiester bond

• In case of no 3’OH no it cannot form a phosphodiester bond

• CHAIN TERMINATION

SEQUENCING USING BACTERIOPHAGE M13

BY PRIMER WALKING

• Molecular Biology by David P Clark

• Cell and Molecular Biology (6th edition) by Gerald Karp

REFERENCES

THANK YOU

High-throughput DNA Sequencing in Microbial Ecology · Luisa Warchavchik Hugerth (2016): High-throughput DNA Sequencing in Microbial Ecology: methods and applications. Division of

DNA sequencing - ULisboabmg.fc.ul.pt/Disciplinas/GBM/aulas/5DNAsequencing.pdf · DNA sequencing methods with fluorophores. 35S -(a -thio) -deoxynucleoside triphosphate ... a laser

DNA SEQUENCING METHODS

DNA sequencing: methods I. Brief history of sequencing II. Sanger dideoxy method for sequencing III. Sequencing large pieces of DNA VI. The “$1,000 dollar

Sanger Sequencing - KSU · •The term DNA sequencing refers to ….. •Application? •A sequencing can be done by different methods including: 1. Maxam–Gilbert sequencing (chemical

Sequencing Neanderthal DNA Lauren Edelson. Road Map Sequencing ancient DNA: methods & outcomes (Review article) DNA sequencing to infer Neanderthal ancestry

DNA sequencing methods - Open University of Sri Lanka · Sequencing large pieces of DNA: the “shotgun” method • Break DNA into small pieces (typically sizes of around 1000 base

DNA Sequencing, Bioinformatics and Microarrays. DNA Sequencing Today, laboratories routinely sequence the order of nucleotides in DNA. DNA sequencing

Sanger Sequencing - KSU · DNA sequencing: • The term DNA sequencing refers to ….. •A sequencing can be done by different methods including: 1. Maxam –Gilbert sequencing (chemical

[PPT]Methods in DNA Sequencing - University of Texas at Austintandy/zach.ppt · Web viewMethods in DNA Sequencing Big Picture Large-scale sequencing requires DNA to be broken into

Dna Sequencing

Dna sequencing

Genome Characterization DNA sequence-ULTIMATE Map DNA sequencing-methods Assembly/sequencing BIO520 BioinformaticsJim Lund Assigned reading: Service 2006

BMC Microbiology BioMed Central...tions based on molecular methods such as DNA-DNA-hybridization, 16S rRNA sequencing, PCR-based methods with primers derived from rRNA sequencing,

Automated DNA Sequencing - UAlbertaCycle sequencing requires much less template DNA than single-temperature extension methods. Cycle sequencing is more convenient than traditional

DNA Sequencing: Current State and Prospects of Development · 2019-07-16 · DNA Sequencing Methods DNA sequencing methods are: Basic DNA sequencing • Sanger method (chain termination)

Different methods of gene isolation, Techniques of DNA sequencing, Artificial … · 2018-03-14 · Different methods of gene isolation, Techniques of DNA sequencing, Artificial DNA

DNA sequencing - Bilkent University Sequencing: History Sanger method (1977): labeled ddNTPs terminate DNA copying at random points. Both methods generate labeled fragments of

1Sources of DNA and Sequencing Methods.1Sources of DNA and Sequencing Methods 2 Genome Assembly Strategy and Characterization 2 Genome Assembly

DNA Sequencing Methods Past, Present, and Futureindico.ictp.it/event/a10159/session/8/contribution/7/material/0/0.pdf · DNA Sequencing Methods Past, Present, and Future Author: Arjang

DNA sequencing methods. Developing new methods and instruments that permit fast polynucleotide sequencing has attracted considerable attention recently

Methods for Working with DNA and RNA - siumed.edubbartholomew/-lectures/DNA methods pt2 '08.pdf · Methods for Working with DNA and RNA 4. DNA sequencing ... Methods for Working with

Next-Generation DNA Sequencing Methods

DNA sequencing - Caldwell-West Caldwell Public Schools...3 Sequencing DNA: •Rapid DNA sequencing methods were first developed in the mid 1970's. •DNA sequencing has developed rapidly;

Methods of DNA Sequencing