Sanger Sequencing By D Gnanasingh Arputhadas
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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.
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Next generation sequencing
This document provides an overview of DNA sequencing technologies. It begins with a brief history of DNA sequencing, including the discovery of DNA's structure and Sanger sequencing. The document then focuses on next generation sequencing technologies, describing several platforms such as 454 sequencing, Illumina sequencing, Ion Torrent sequencing, and Pacific Biosciences sequencing. It also discusses third generation sequencing and compares the sequencing approaches, workflows, and applications of various sequencing technologies. In conclusion, the document notes the progress and future directions of sequencing, including increased clinical applications and reduced costs.
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This document discusses DNA sequencing methods, both current and developing technologies. It begins by explaining Sanger sequencing and how fluorescent dyes and thermal cycling improved it. High-throughput short and long-read sequencing methods are then outlined, including Illumina, Ion Torrent, Nanopore, and SMRT sequencing. Developing methods like tunneling currents, hybridization, and microscopy techniques are also mentioned. Overall, the document provides a comprehensive overview of the major DNA sequencing techniques used today and those under investigation.
Principle and workflow of whole genome bisulfite sequencing
Whole genome bisulfite sequencing (WGBS) provides genome-wide single-base resolution DNA methylation profiling. It works by converting unmethylated cytosines to uracils through bisulfite treatment, followed by sequencing and comparing cytosines and thymines to determine methylation status. The workflow involves DNA extraction, bisulfite conversion, library preparation, sequencing, and bioinformatics analysis. WGBS allows for high resolution methylation analysis but has challenges with alignment due to bisulfite conversion.
Molecular structure of genes and chromosomes
The document discusses the molecular structure of genes and chromosomes. It describes how DNA is organized into chromosomes, which contain both protein-coding genes and non-coding sequences. Genes contain exons and introns, and in eukaryotes genes are further organized into transcription units. Chromatin compacts the DNA into nucleosomes and higher-order structures like the 30nm fiber. Overall the document provides an overview of the molecular organization and components that make up eukaryotic genes and chromosomes.
Sanger sequencing
DNA sequencing is the process of determining the order of nucleotides in a DNA molecule. The Sanger method, developed in 1977, was the most widely used sequencing technique for 25 years. It utilizes chain termination with dideoxynucleotides which lack a 3' OH group, preventing formation of a phosphodiester bond and terminating strand elongation. Four reactions are run in parallel with each dideoxynucleotide labeled with a different color. Gel electrophoresis separates the terminated fragments by size, allowing the DNA sequence to be read by matching fragment sizes to nucleotide colors.
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This document discusses pseudogenes, which are dysfunctional copies of genes that have lost protein-coding ability. It covers the origin and formation of pseudogenes through DNA or RNA duplication, and describes different types like processed and unprocessed pseudogenes. The document also discusses various methods for identifying and detecting pseudogenes, databases of pseudogenes, and studies that have characterized pseudogenes in organisms like rice and Solanum plants. Finally, it explores the potential functions and utilities of pseudogenes, including their use in evolutionary studies, providing information about gene expression, and acting as competing endogenous RNAs.
Sanger sequencing
A presentation of how to sequence DNA. Basics of Sanger Sequencing. All animations are created by me. Photographs and graffiti is taken from internet.
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This document provides an overview of DNA sequencing technologies. It begins with a brief history of DNA sequencing, including the discovery of DNA's structure and Sanger sequencing. The document then focuses on next generation sequencing technologies, describing several platforms such as 454 sequencing, Illumina sequencing, Ion Torrent sequencing, and Pacific Biosciences sequencing. It also discusses third generation sequencing and compares the sequencing approaches, workflows, and applications of various sequencing technologies. In conclusion, the document notes the progress and future directions of sequencing, including increased clinical applications and reduced costs.
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This document discusses DNA sequencing methods, both current and developing technologies. It begins by explaining Sanger sequencing and how fluorescent dyes and thermal cycling improved it. High-throughput short and long-read sequencing methods are then outlined, including Illumina, Ion Torrent, Nanopore, and SMRT sequencing. Developing methods like tunneling currents, hybridization, and microscopy techniques are also mentioned. Overall, the document provides a comprehensive overview of the major DNA sequencing techniques used today and those under investigation.
Principle and workflow of whole genome bisulfite sequencing
Whole genome bisulfite sequencing (WGBS) provides genome-wide single-base resolution DNA methylation profiling. It works by converting unmethylated cytosines to uracils through bisulfite treatment, followed by sequencing and comparing cytosines and thymines to determine methylation status. The workflow involves DNA extraction, bisulfite conversion, library preparation, sequencing, and bioinformatics analysis. WGBS allows for high resolution methylation analysis but has challenges with alignment due to bisulfite conversion.
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The document discusses the molecular structure of genes and chromosomes. It describes how DNA is organized into chromosomes, which contain both protein-coding genes and non-coding sequences. Genes contain exons and introns, and in eukaryotes genes are further organized into transcription units. Chromatin compacts the DNA into nucleosomes and higher-order structures like the 30nm fiber. Overall the document provides an overview of the molecular organization and components that make up eukaryotic genes and chromosomes.
Sanger sequencing
DNA sequencing is the process of determining the order of nucleotides in a DNA molecule. The Sanger method, developed in 1977, was the most widely used sequencing technique for 25 years. It utilizes chain termination with dideoxynucleotides which lack a 3' OH group, preventing formation of a phosphodiester bond and terminating strand elongation. Four reactions are run in parallel with each dideoxynucleotide labeled with a different color. Gel electrophoresis separates the terminated fragments by size, allowing the DNA sequence to be read by matching fragment sizes to nucleotide colors.
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This powerpoint explains about the nucleic acid hybridization, its principle, application and the assay methods. Also it gives clear picture about DNA probes, its sysnthesis, mechanism of probes and the detector system in DNA hybridization.
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This document discusses pseudogenes, which are dysfunctional copies of genes that have lost protein-coding ability. It covers the origin and formation of pseudogenes through DNA or RNA duplication, and describes different types like processed and unprocessed pseudogenes. The document also discusses various methods for identifying and detecting pseudogenes, databases of pseudogenes, and studies that have characterized pseudogenes in organisms like rice and Solanum plants. Finally, it explores the potential functions and utilities of pseudogenes, including their use in evolutionary studies, providing information about gene expression, and acting as competing endogenous RNAs.
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A presentation of how to sequence DNA. Basics of Sanger Sequencing. All animations are created by me. Photographs and graffiti is taken from internet.
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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.
Next Generation Sequencing
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.
Physical mapping
A physical map of a chromosome or a genome that shows the physical locations of genes and other DNA sequences of interest. Physical maps are used to help scientists identify and isolate genes by positional cloning.
According to the ICSM (Intergovernmental Committee on Surveying and Mapping), there are five different types of maps: General Reference, Topographical, Thematic, Navigation Charts and Cadastral Maps and Plans.
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Mitochondrial DNA (mtDNA) is small, circular, double-stranded DNA located in cell mitochondria. It is maternally inherited and does not recombine. mtDNA contains 37 genes essential for mitochondrial function and ATP production through oxidative phosphorylation. Compared to nuclear DNA, mtDNA evolves more rapidly, lacks introns, and is not bound in histones. Forensic analysis of mtDNA is useful when evidence is degraded or limited. Methods include DNA extraction, PCR amplification of mtDNA regions, sequencing, and comparing sequences to identify matches or mismatches. mtDNA analysis has applications in fisheries including individual identification, mixed stock analysis, and determining phylogenetic relationships between fish species.
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DNA SEQUENCE ANALYSIS.pdf
DNA is the molecule that contains the genetic instructions used in the development and functioning of all known living organisms. A gene is a unit of heredity and consists of a segment of DNA that codes for a protein or RNA molecule. DNA sequencing is the process of determining the precise order of nucleotides within a DNA molecule, and includes any method used to determine the order of the four bases - adenine, thymine, guanine, and cytosine. There are two main historical methods for DNA sequencing - the Maxam-Gilbert chemical method and the Sanger dideoxy chain termination method, upon which modern automated sequencing is based using fluorescence detection. DNA sequencing has many applications in fields like medicine, forensics, and agriculture
Comparative genomic hybridization
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This document defines DNA sequencing and describes some common DNA sequencing methods. It explains that DNA sequencing determines the order of the four nucleotide bases that make up DNA. It then describes two basic DNA sequencing methods - Maxam-Gilbert chemical sequencing and Sanger chain termination sequencing. For Sanger sequencing, it provides details on how fluorescent dideoxynucleotides are used to randomly terminate DNA strands during replication, allowing the sequence to be read from the resulting fragments.
Dna footprinting
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Rflp technology
RFLP is a technique that differentiates organisms by analyzing patterns in DNA fragments produced after digestion with restriction enzymes. If two organisms differ in the distance between restriction sites, the lengths of fragments produced will differ. These patterns can differentiate species and strains. RFLP detection relies on comparing band profiles after digestion and gel electrophoresis to see length polymorphisms, which can then be examined further through hybridization and visualization. RFLPs have applications in forensics, disease detection, and human population genetics.
Lecture on DNA sequencing
The document discusses different methods of DNA sequencing including the Maxam-Gilbert and Sanger chain termination methods as well as newer next generation sequencing techniques. It describes the principles, steps, and significance of the Maxam-Gilbert and Sanger methods and how next generation sequencing improved DNA sequencing by allowing millions of DNA molecules to be sequenced simultaneously in an automated process.
molecular marker RFLP, and application
INTRODUCTION
RFLP
Analysis technique
APPLICATION OF RFLP
FORENSIC
DISEASE PROGNOSIS
GENETIC COUNSELING
PEDIGREE
VARIETAL
Advantages of RFLP
Disadvantages of RFLP
CONCLUSIONS
REFERENCES
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This document discusses DNA sequencing methods. It describes the Maxam-Gilbert sequencing method developed in 1976-1977 which uses chemical modification and cleavage of DNA at specific bases, followed by electrophoresis to separate fragments by size. It also mentions the popular Sanger sequencing method. The procedure for Maxam-Gilbert sequencing involves labeling DNA, cleaving it with chemicals, running the fragments on a gel, and analyzing the results to deduce the DNA sequence. Advantages include no premature termination and ability to sequence stretches not possible with enzymatic methods, while disadvantages include use of radioactivity and toxic chemicals.
Next generation sequencing
Next generation sequencing techniques allow for high-throughput DNA sequencing at a lower cost compared to Sanger sequencing. The document focuses on Illumina sequencing and 454 pyrosequencing. In Illumina sequencing, DNA fragments are attached to a flow cell and undergo bridge amplification and sequencing by synthesis using fluorescently labeled nucleotides. 454 pyrosequencing involves emulsion PCR to amplify DNA fragments attached to beads, followed by sequencing using DNA polymerase and a bioluminescent detection of incorporated nucleotides. Both techniques allow for massively parallel sequencing of millions of DNA fragments.
dna sequencing methods
1. DNA sequencing involves determining the order of nucleotide bases in DNA. The original chain termination or dideoxy method developed by Sanger is still widely used for small DNA segments.
2. Whole genome shotgun sequencing breaks large genomes into fragments that are sequenced and then reassembled, allowing sequencing of entire genomes.
3. Pyrosequencing is a sequencing by synthesis method that uses a bioluminescent reaction to determine nucleotides added, enabling accurate and fast sequencing.
Dna Footprinting
DNA footprinting is a technique used to identify where proteins bind to DNA. It was developed in 1978 and works by treating DNA with enzymes or chemicals that cut DNA, except for regions bound by proteins. This leaves a "footprint" where the protein is bound and protects the DNA. There are two main types: DNase I footprinting cuts DNA randomly except where proteins are bound, while DMS footprinting modifies DNA bases except where proteins protect them from modification. The cut or modified DNA is then run on a gel to identify the protein binding site. DNA footprinting is useful for mapping transcription factor binding sites and studying protein-DNA interactions.
DNA sequencing- Maxam- Gilbert sequencing
published a DNA sequencing method in 1977 based on chemical modification of DNA and subsequent cleavage at specific bases. Also known as chemical sequencing, this method allowed purified samples of double-stranded DNA to be used without further cloning.
Maxam-Gilbert sequencing requires radioactive labeling at one 5' end of the DNA and purification of the DNA fragment to be sequenced. Chemical treatment then generates breaks at a small proportion of one or two of the four nucleotide bases in each of four reactions (G, A+G, C, C+T). The concentration of the modifying chemicals is controlled to introduce on average one modification per DNA molecule. Thus a series of labeled fragments is generated, from the radiolabeled end to the first "cut" site in each molecule. The fragments in the four reactions are electrophoresed side by side in denaturing acrylamide gels for size separation. To visualize the fragments, the gel is exposed to X-ray film for autoradiography, yielding a series of dark bands each corresponding to a radiolabeled DNA fragment, from which the sequence may be inferred.
RFLP
This document discusses Restriction Fragment Length Polymorphism (RFLP) analysis, which is a technique used to differentiate organisms by analyzing patterns in their DNA after digestion with restriction enzymes. RFLP analysis can be used for various applications like determining paternity, detecting mutations, and genetic mapping. The process involves digesting DNA with restriction enzymes, running the fragments on a gel, transferring the DNA to a membrane, and using probes to detect polymorphisms and produce an autoradiogram showing differences in fragment patterns. As an example, the document describes using PCR-RFLP to rapidly screen for a BRCA2 mutation by taking advantage of a restriction site change caused by the mutation.
Sanger sequencing
The document discusses Sanger sequencing, a method of DNA sequencing. It provides a brief history of DNA sequencing, noting that Sanger developed an enzymatic DNA sequencing technique in 1977. The document then describes the key steps of Sanger sequencing, including separating the DNA strands, copying one strand with chemically altered bases that cause termination, and analyzing the fragments to reveal the DNA sequence. It also compares Sanger sequencing to the Maxam-Gilbert chemical degradation method.
Dna Sequencing
The document summarizes DNA sequencing methods. It discusses the DNA double helix structure and how the four nitrogenous bases form complementary pairs between strands. It then describes the two main historical DNA sequencing methods: the Maxam-Gilbert method which uses chemical degradation, and the Sanger method which is based on chain termination using dideoxynucleotides. The Sanger method is now the most common approach and involves sequencing in four separate reactions with one of the four ddNTPs added to each.
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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.
Next Generation Sequencing
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.
Physical mapping
A physical map of a chromosome or a genome that shows the physical locations of genes and other DNA sequences of interest. Physical maps are used to help scientists identify and isolate genes by positional cloning.
According to the ICSM (Intergovernmental Committee on Surveying and Mapping), there are five different types of maps: General Reference, Topographical, Thematic, Navigation Charts and Cadastral Maps and Plans.
NGS data formats and analyses
Presentation to cover the data and file formats commonly used in next generation sequencing (high throughput sequencing) analyses. From nucleotide ambiguity codes, FASTA and FASTQ, quality scores to SAM and BAM, CIGAR strings and variant calling format. This was given as part of the EPIZONE Workshop on Next Generation Sequencing applications and Bioinformatics in Brussels, Belgium in April 2016.
Mt DNA
Mitochondrial DNA (mtDNA) is small, circular, double-stranded DNA located in cell mitochondria. It is maternally inherited and does not recombine. mtDNA contains 37 genes essential for mitochondrial function and ATP production through oxidative phosphorylation. Compared to nuclear DNA, mtDNA evolves more rapidly, lacks introns, and is not bound in histones. Forensic analysis of mtDNA is useful when evidence is degraded or limited. Methods include DNA extraction, PCR amplification of mtDNA regions, sequencing, and comparing sequences to identify matches or mismatches. mtDNA analysis has applications in fisheries including individual identification, mixed stock analysis, and determining phylogenetic relationships between fish species.
Polymerase Chain Reaction
The Polymerase Chain Reaction (PCR) is a technique that allows for the amplification of specific DNA sequences. It involves cycling between heating and cooling steps to denature, anneal primers to, and extend DNA. This allows a small amount of DNA to be exponentially replicated, enabling applications like disease diagnosis, genetic identification, and DNA analysis. PCR requires DNA, primers, DNA polymerase, nucleotides, and thermal cycling to replicate the target DNA sequence.
Dna Fingerprinting And Forensic Applications
DNA fingerprinting is a technique used to identify individuals by their unique DNA patterns. It involves extracting DNA from samples, cutting the DNA into fragments using restriction enzymes, separating the fragments via electrophoresis, and comparing the band patterns to determine if two DNA samples match. DNA fingerprinting is used in paternity testing, criminal identification, and other forensic applications to compare DNA evidence from a crime scene to a suspect's DNA.
DNA SEQUENCE ANALYSIS.pdf
DNA is the molecule that contains the genetic instructions used in the development and functioning of all known living organisms. A gene is a unit of heredity and consists of a segment of DNA that codes for a protein or RNA molecule. DNA sequencing is the process of determining the precise order of nucleotides within a DNA molecule, and includes any method used to determine the order of the four bases - adenine, thymine, guanine, and cytosine. There are two main historical methods for DNA sequencing - the Maxam-Gilbert chemical method and the Sanger dideoxy chain termination method, upon which modern automated sequencing is based using fluorescence detection. DNA sequencing has many applications in fields like medicine, forensics, and agriculture
Comparative genomic hybridization
Comparative genomic hybridization (CGH) is a molecular cytogenetic technique that allows detection of copy number variations between a test and reference DNA sample without cell culturing. CGH involves labeling and hybridizing test and reference DNA to normal metaphase chromosomes before visualizing differences in fluorescence to identify regions of gains or losses. While CGH was originally used for cancer research, it can also detect chromosomal abnormalities associated with genetic disorders and has improved resolution over traditional cytogenetic methods. The main limitations of CGH are its inability to detect structural aberrations without copy number changes and resolutions above 5-10 megabases.
DND sequencing
This document defines DNA sequencing and describes some common DNA sequencing methods. It explains that DNA sequencing determines the order of the four nucleotide bases that make up DNA. It then describes two basic DNA sequencing methods - Maxam-Gilbert chemical sequencing and Sanger chain termination sequencing. For Sanger sequencing, it provides details on how fluorescent dideoxynucleotides are used to randomly terminate DNA strands during replication, allowing the sequence to be read from the resulting fragments.
Dna footprinting
DNA footprinting is a method of investigating the sequence specificity of DNA-binding proteins in vitro.
Rflp technology
RFLP is a technique that differentiates organisms by analyzing patterns in DNA fragments produced after digestion with restriction enzymes. If two organisms differ in the distance between restriction sites, the lengths of fragments produced will differ. These patterns can differentiate species and strains. RFLP detection relies on comparing band profiles after digestion and gel electrophoresis to see length polymorphisms, which can then be examined further through hybridization and visualization. RFLPs have applications in forensics, disease detection, and human population genetics.
Lecture on DNA sequencing
The document discusses different methods of DNA sequencing including the Maxam-Gilbert and Sanger chain termination methods as well as newer next generation sequencing techniques. It describes the principles, steps, and significance of the Maxam-Gilbert and Sanger methods and how next generation sequencing improved DNA sequencing by allowing millions of DNA molecules to be sequenced simultaneously in an automated process.
molecular marker RFLP, and application
INTRODUCTION
RFLP
Analysis technique
APPLICATION OF RFLP
FORENSIC
DISEASE PROGNOSIS
GENETIC COUNSELING
PEDIGREE
VARIETAL
Advantages of RFLP
Disadvantages of RFLP
CONCLUSIONS
REFERENCES
Maxam–Gilbert sequencing
This document discusses DNA sequencing methods. It describes the Maxam-Gilbert sequencing method developed in 1976-1977 which uses chemical modification and cleavage of DNA at specific bases, followed by electrophoresis to separate fragments by size. It also mentions the popular Sanger sequencing method. The procedure for Maxam-Gilbert sequencing involves labeling DNA, cleaving it with chemicals, running the fragments on a gel, and analyzing the results to deduce the DNA sequence. Advantages include no premature termination and ability to sequence stretches not possible with enzymatic methods, while disadvantages include use of radioactivity and toxic chemicals.
Next generation sequencing
Next generation sequencing techniques allow for high-throughput DNA sequencing at a lower cost compared to Sanger sequencing. The document focuses on Illumina sequencing and 454 pyrosequencing. In Illumina sequencing, DNA fragments are attached to a flow cell and undergo bridge amplification and sequencing by synthesis using fluorescently labeled nucleotides. 454 pyrosequencing involves emulsion PCR to amplify DNA fragments attached to beads, followed by sequencing using DNA polymerase and a bioluminescent detection of incorporated nucleotides. Both techniques allow for massively parallel sequencing of millions of DNA fragments.
dna sequencing methods
1. DNA sequencing involves determining the order of nucleotide bases in DNA. The original chain termination or dideoxy method developed by Sanger is still widely used for small DNA segments.
2. Whole genome shotgun sequencing breaks large genomes into fragments that are sequenced and then reassembled, allowing sequencing of entire genomes.
3. Pyrosequencing is a sequencing by synthesis method that uses a bioluminescent reaction to determine nucleotides added, enabling accurate and fast sequencing.
Dna Footprinting
DNA footprinting is a technique used to identify where proteins bind to DNA. It was developed in 1978 and works by treating DNA with enzymes or chemicals that cut DNA, except for regions bound by proteins. This leaves a "footprint" where the protein is bound and protects the DNA. There are two main types: DNase I footprinting cuts DNA randomly except where proteins are bound, while DMS footprinting modifies DNA bases except where proteins protect them from modification. The cut or modified DNA is then run on a gel to identify the protein binding site. DNA footprinting is useful for mapping transcription factor binding sites and studying protein-DNA interactions.
DNA sequencing- Maxam- Gilbert sequencing
published a DNA sequencing method in 1977 based on chemical modification of DNA and subsequent cleavage at specific bases. Also known as chemical sequencing, this method allowed purified samples of double-stranded DNA to be used without further cloning.
Maxam-Gilbert sequencing requires radioactive labeling at one 5' end of the DNA and purification of the DNA fragment to be sequenced. Chemical treatment then generates breaks at a small proportion of one or two of the four nucleotide bases in each of four reactions (G, A+G, C, C+T). The concentration of the modifying chemicals is controlled to introduce on average one modification per DNA molecule. Thus a series of labeled fragments is generated, from the radiolabeled end to the first "cut" site in each molecule. The fragments in the four reactions are electrophoresed side by side in denaturing acrylamide gels for size separation. To visualize the fragments, the gel is exposed to X-ray film for autoradiography, yielding a series of dark bands each corresponding to a radiolabeled DNA fragment, from which the sequence may be inferred.
RFLP
This document discusses Restriction Fragment Length Polymorphism (RFLP) analysis, which is a technique used to differentiate organisms by analyzing patterns in their DNA after digestion with restriction enzymes. RFLP analysis can be used for various applications like determining paternity, detecting mutations, and genetic mapping. The process involves digesting DNA with restriction enzymes, running the fragments on a gel, transferring the DNA to a membrane, and using probes to detect polymorphisms and produce an autoradiogram showing differences in fragment patterns. As an example, the document describes using PCR-RFLP to rapidly screen for a BRCA2 mutation by taking advantage of a restriction site change caused by the mutation.
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Dna Sequencing
The document summarizes DNA sequencing methods. It discusses the DNA double helix structure and how the four nitrogenous bases form complementary pairs between strands. It then describes the two main historical DNA sequencing methods: the Maxam-Gilbert method which uses chemical degradation, and the Sanger method which is based on chain termination using dideoxynucleotides. The Sanger method is now the most common approach and involves sequencing in four separate reactions with one of the four ddNTPs added to each.
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This document discusses the history and various methods of DNA sequencing. It begins with a brief overview of DNA sequencing and its uses. It then outlines some of the major developments in DNA sequencing techniques, including the earliest RNA sequencing in 1972, Sanger sequencing in 1977, and the first complete genome of Haemophilus influenzae in 1995. The document proceeds to provide more detailed explanations of several DNA sequencing methods, such as Sanger sequencing, pyrosequencing, shotgun sequencing, Illumina sequencing, and SOLiD sequencing.
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DNA sequencing determines the order of nucleotide bases in a DNA molecule. The first methods were developed in the 1970s by Maxam and Gilbert (chemical method) and Sanger (chain termination/dideoxy method). Sanger's method is now most commonly used and involves DNA synthesis with chain termination by dideoxynucleotides to generate fragments of different lengths that can be separated and read to determine the DNA sequence. DNA sequencing has revolutionized biological sciences by enabling diagnosis of genetic diseases, identification of disease-causing mutations, and mapping of genomes. It provides benefits for medicine, forensics, and agriculture.
2013 CSM
Student Symposium talk given at the Canadian Society of Microbiologist's meeting in 2013. Won the ISME 8 Student Symposium Award.
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This document describes the process of Southern blotting, which is used to detect specific DNA sequences in a sample. It involves purifying DNA from cells, cutting the DNA into fragments using restriction enzymes, separating the fragments by size via gel electrophoresis, transferring them to a membrane, then using a radioactive probe to detect fragments that hybridize based on complementary base pairing. The technique allows identification of mutations, deletions and other genetic variations, and has applications in medical diagnosis, forensics and other areas requiring DNA analysis.
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Similar to Sanger Sequencing By D Gnanasingh Arputhadas
DNA Sequencing
This document provides an overview of DNA sequencing:
- It discusses the history of DNA sequencing, from the early 1970s methods to modern techniques. The Sanger and Maxam-Gilbert methods were among the first developed.
- DNA sequencing involves determining the order of nucleotides (A, T, C, G) in a DNA molecule. This provides important information for research and applications in diagnostics, biotechnology, forensics, and more.
- The document outlines some of the major DNA sequencing techniques and methods, including Sanger sequencing and Maxam-Gilbert sequencing. It also discusses next-generation sequencing approaches.
DNA SEQUENCING by sanger method.pptx
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DNA SEQUENCING Shashi kala.prasentation
The document discusses DNA sequencing and its importance. It begins with defining DNA and describing its structure and discovery. It then explains how DNA sequencing works, including the Sanger sequencing method. The key points made are:
1. DNA sequencing determines the order of nucleotides in DNA and has helped accelerate biological research.
2. Sanger sequencing uses dideoxynucleotides to terminate DNA synthesis at different positions, producing fragments of different lengths that can be read to determine the sequence.
3. Knowing DNA sequences helps determine protein functions, aids medical diagnosis and research, and allows comparison of healthy and mutated DNA. This knowledge is important for identifying diseases like cancer and viruses.
Dna sequencing
The document summarizes a presentation on DNA sequencing techniques. It describes DNA and its functions of storing genetic information, self-duplication, and expressing genetic messages. It then explains the historical Maxam-Gilbert and Sanger DNA sequencing methods and how modern sequencing uses variations of the Sanger technique. The document provides details on the chemical degradation approach of Maxam-Gilbert and the chain termination approach of Sanger sequencing before polymerase chain reaction and gel electrophoresis. It compares the two methods and discusses applications and future directions of DNA sequencing.
Presentation1
This document discusses DNA sequencing, including its history, different methods, principles, requirements, procedures, importance, purposes, and applications. It describes two main DNA sequencing methods - Maxam-Gilbert sequencing and Sanger sequencing. Maxam-Gilbert sequencing uses chemical treatment to generate breaks in DNA at specific bases, while Sanger sequencing uses DNA polymerase and dideoxynucleotides to terminate DNA strand extension. The document also outlines how DNA sequencing is used in fields like forensics, medicine, and agriculture.
-DNA Sequencing Notes.pdf
DNA sequencing is a laboratory technique used to determine the exact sequence of bases (A, C, G, and T) in a DNA molecule. The DNA base sequence carries the information a cell needs to assemble protein and RNA molecules. DNA sequence information is important to scientists investigating the functions of genes.
In medicine, DNA sequencing is used for a range of purposes, including diagnosis and treatment of diseases. In general, sequencing allows health care practitioners to determine if a gene or the region that regulates a gene contains changes, called variants or mutations, that are linked to a disorder.
-DNA Sequencing Notes.pdf
DNA sequencing refers to the general laboratory technique for determining the exact sequence of nucleotides, or bases, in a DNA molecule. The sequence of the bases (often referred to by the first letters of their chemical names: A, T, C, and G) encodes the biological information that cells use to develop and operate. Establishing the sequence of DNA is key to understanding the function of genes and other parts of the genome. There are now several different methods available for DNA sequencing, each with its own characteristics, and the development of additional methods represents an active area of genomics research.
DNA sequencing
Genome sequencing and comparative genomics are important tools in plant breeding. Genome sequencing determines the order of DNA nucleotides in individual genes, chromosomes, or entire genomes. Comparative genomics analyzes and compares genetic material between species to study evolution, gene function, and disease. Next generation sequencing techniques like Illumina sequencing have made genome sequencing faster, cheaper, and able to sequence thousands of sequences at once. Comparative genomics is used to understand differences between species by comparing gene location, structure, sequence similarity and other characteristics. This aids in understanding evolution and identifying genes responsible for unique traits.
Dna sequencing ppt
This document discusses DNA sequencing methods and their history and applications. It covers first generation sequencing methods like Sanger sequencing and Maxam-Gilbert sequencing. It also covers next generation sequencing (NGS) methods like 454 pyrosequencing, Illumina sequencing, and Ion Torrent semiconductor sequencing. NGS allows high-throughput, massively parallel sequencing of DNA fragments. Template preparation for NGS involves fragmenting DNA, attaching fragments to beads, and emulsion PCR. The document provides details on the chemistry and detection methods used for different sequencing platforms.
Dna sequencing
The document discusses DNA sequencing and its applications. It describes DNA sequencing as determining the exact order of A, T, C, and G bases that make up human chromosomes. The Sanger dideoxy method was developed in 1977 and remains a popular sequencing technique that uses DNA replication with a primer, polymerase, and tagged nucleotides. DNA sequencing has applications in forensics, medicine, and agriculture such as identifying individuals, detecting genes linked to disorders, and improving crops.
DNA sequencing methods
DNA SEQUENCING:
MAXAM GILBERT METHOD
SANGER SEQUENCING
Illumina (Solexa) sequencing
Pyrosequencing
Presentation on dna sequencing
This document provides an overview of DNA sequencing. It begins by defining DNA and its structure. It then explains that DNA sequencing is the process of determining the order of nucleotide bases in a DNA strand. Two common methods of DNA sequencing are described in detail: the Maxam-Gilbert chemical cleavage method and the Sanger dideoxy chain termination method. The document concludes by discussing some applications of DNA sequencing in fields like forensics, medical research, and agriculture.
Bahaddin DNA sequencing
The document provides information on several DNA sequencing methods including:
- The Maxam-Gilbert method which uses chemical cleavage to sequence DNA.
- The Sanger method (chain-termination method) which uses dideoxynucleotides and DNA polymerase to terminate DNA strand extension at specific bases.
- Next generation sequencing methods like 454 pyrosequencing, Illumina sequencing, and Ion Torrent sequencing that allow for massively parallel sequencing of many DNA fragments simultaneously.
The history and basic principles of the Maxam-Gilbert and Sanger methods are described in detail as they were the first widely used DNA sequencing techniques. Subsequent methods aimed to improve speed, throughput and reduce cost of sequencing
DNA Sequencing
The document discusses DNA sequencing, including its history, common methods like Sanger sequencing, the steps involved, challenges and limitations. DNA sequencing determines the order of nucleotides in DNA and has various applications such as identifying genetic diseases, forensics, and studying genome evolution. Sanger sequencing, developed in 1977, works by using chain-terminating dideoxynucleotides during DNA replication to determine the sequence.
Dna sequencing.
DNA sequencing refers to determining the order of nucleotide bases in a DNA molecule. Frederick Sanger developed the chain termination/dideoxy method in 1977, which uses DNA polymerase and dideoxynucleotides to generate fragmented DNA strands of different lengths. Pyrosequencing was later developed, detecting light pulses from nucleotide incorporation without electrophoresis. Both methods have advanced biological research and applications in medicine, biotechnology and forensics.
genesequencing-200105073623 (1).pdf
Gene sequencing is the technique that determines the order of nucleotide bases in DNA. It allows researchers to read genetic information and understand genes. The first genome sequenced was a bacteriophage in 1977. Major advances include sequencing the human genome in 2001. Current technologies like Illumina and Ion Torrent can generate billions of reads faster and cheaper than Sanger sequencing. Gene sequencing has applications in medicine, forensics, agriculture, and more. It is an important tool for understanding genomes and their relationship to traits and disease.
Gene Sequencing
Gene sequencing is the technique that determines the order of nucleotide bases in DNA. It allows researchers to read genetic information and understand genes. The first genome sequenced was a bacteriophage in 1977. Techniques have advanced from Sanger sequencing to second-generation sequencing using platforms like Illumina and third-generation single-molecule techniques. Gene sequencing has various applications in medicine, forensics, agriculture, cancer research and more. It is an important tool for understanding genomes and their relationship to traits and disease.
DNA Function and Structure Notes
DNA contains all of an organism's genetic information and is found in the cells of all living things. DNA is made up of long chains of nucleotides, which consist of a sugar, phosphate, and one of four nitrogen-containing bases. The order of these bases in the DNA determines an organism's traits by encoding genes. James Watson and Francis Crick discovered that DNA exists as a double helix structure, with the bases pairing together in a complementary way between chains.
SIR WASIQ dna sequence ppt.pptx
1) DNA sequencing determines the order of nucleotides (A, T, C, G) in DNA that provide genetic information. A pairs with T and C pairs with G.
2) Sanger sequencing, discovered in the 1970s, was the first DNA sequencing method and uses fluorescent ddNTPs to terminate nucleotide addition. Next-generation sequencing now allows millions of fragments to be sequenced simultaneously.
3) DNA sequencing reveals genetic information and can identify genes, compare homologous sequences between species, and detect changes causing disease. It has applications in medicine, agriculture, food safety, and environmental protection.
sequencing.pptx
DNA sequencing is the process of determining the order of nucleotides in DNA. Friedrich Miescher first discovered DNA in 1869. In 1977, Frederick Sanger developed the chain-termination method, which is still used today. DNA sequencing reveals the genetic information carried in a DNA segment and has applications in medicine, evolutionary analysis, disease diagnosis, food safety and more. It works by separating single-stranded DNA molecules that differ by a single nucleotide using gel electrophoresis.
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Sanger Sequencing By D Gnanasingh Arputhadas
- 2. DNA (Deoxyribonucleic acid ) Deoxyribonucleic acid (DNA) is a molecule that encodes the genetic instructions used in the development and functioning of all known living organisms and many viruses. DNA is a nucleic acid; alongside proteins and carbohydrates, nucleic acids compose the three major macromolecules essential for all known forms of life. Two Strands Double Helix
- 3. DNA Sequencing DNA sequencing is the process of determining the precise order of nucleotides within a DNA molecule. It includes any method or technology that is used to determine the order of the four bases—adenine, guanine, cytosine, and thymine—in a strand of DNA. The first DNA sequences were obtained in the early 1970s by academic researchers using laborious methods based on two-dimensional chromatography.
- 4. Basic Methods Maxam-Gilbert sequencing Chain-termination methods (Sanger sequencing)
- 5. Sanger sequencing Sanger sequencing is a DNA sequencing method in which target DNA is denatured and annealed to an oligonucleotide primer, which is then extended by DNA polymerase using a mixture of deoxynucleotide triphosphates (normal dNTPs) and chain-terminating dideoxynucleotide triphosphates (ddNTPs).
- 6. Sanger sequencing Developed by Frederick Sanger and colleagues in 1977. It was the most widely used sequencing method for approximately 25 years.
- 7. DNA Sequencing (Video) Video Link : http://www.youtube.com/watch?v=6ldtdWjDwes
- 8. Sanger Sequencing (Complete Process) Video Link : http://www.youtube.com/watch?v=vK-HlMaitnE
- 12. PROCESS
- 13. Triphosphates ATP - Adenosine triphosphate CTP - Cytidine triphosphate TTP - Thiamine triphosphate GTP - Guanosine triphosphate
- 14. RESULT Negative End Positive End
- 15. Need For Sequencing Evolutionary biology DNA fingerprinting Detect the presence of known genes for medical purposes (see genetic testing) Forensic identification Parental testing Detecting mutations
- 16. Applications of DNA Sequencing Forensics Identify individuals Determine the paternity of a child Identifies endangered and protected species Medicine Detect genes that are hereditary or cause diseases Agriculture Map the genome of microorganisms
- 17. Future of DNA Sequencing Projects might focus on researching: The links to develop lifestyle Genomic and cardiovascular disease Early detections of cancer.