Dna sequencing
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The document discusses DNA sequencing and the Sanger method. It explains that the Sanger method involves (1) denaturing double-stranded DNA into single strands, (2) attaching primers and extending DNA fragments with nucleotides except one reaction uses a dideoxynucleotide which terminates extension, (3) separating the fragments by size using gel electrophoresis, and (4) determining the sequence by the termination positions. The Sanger method is commonly used today to determine the sequence of nucleotides in DNA and RNA from various organisms, which has applications in molecular biology, evolutionary biology, and other fields.
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DNA sequencing
DNA sequencing refers to determining the order of nucleotides in a DNA molecule. The first DNA sequence was obtained in the 1970s using chromatography. Modern methods use dye-based sequencing and automation. The two main historical methods are the Maxam-Gilbert chemical degradation method and the Sanger dideoxy chain termination method. Next generation sequencing now allows millions of DNA molecules to be sequenced in parallel through massively parallel sequencing technologies.
DNA sequencing methods
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.
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.
Synthetic Genome
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Dna sequencing
DNA sequencing determines the order of nucleotide bases in a DNA sample. The Sanger method, also known as dideoxy or chain termination method, was the first widely used DNA sequencing technique. It involves DNA polymerase, dNTPs, a primer, and ddNTPs to terminate DNA strand extension. The fragments are separated by gel electrophoresis and the sequence is read. Next-generation sequencing methods like pyrosequencing do not require electrophoresis or fragment separation. They detect incorporated nucleotides in real-time. Nanopore sequencing detects changes in electrical current as single-stranded DNA passes through a nanopore protein.
Construction of genomic library in lambda
Construction of genomic library in lambda phage.
THIS WAS MY PRESENTATION TOPIC IN CLASS. THOUGHT OF SHARING IT AND HOPE IT HELPS.
An introduction to RNA-seq data analysis
AGRF in conjunction with EMBL Australia recently organised a workshop at Monash University Clayton. This workshop was targeted at beginners and biologists who are new to analysing Next-Gen Sequencing data. The workshop also aimed to provide users with a snapshot of bioinformatics and data analysis tips on how to begin to analyse project data. An introduction to RNA-seq data analysis was presented by AGRF Senior Bioinformatician Dr. Sonika Tyagi.
Presented: 1st August 2012
Dna sequencing
DNA sequencing is a process to determine the order of nucleotides in a DNA molecule. It was discovered in the 1970s by scientists like Frederick Sanger who developed the chain termination method. This method involves DNA replication with modified nucleotides that cause the growing DNA strand to terminate at that point. The fragments are then separated by size to reveal the sequence. Automated sequencing now uses fluorescent dyes and capillary electrophoresis for faster and higher throughput sequencing. DNA sequencing has applications in medicine, forensics, and agriculture.
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DNA sequencing
DNA sequencing refers to determining the order of nucleotides in a DNA molecule. The first DNA sequence was obtained in the 1970s using chromatography. Modern methods use dye-based sequencing and automation. The two main historical methods are the Maxam-Gilbert chemical degradation method and the Sanger dideoxy chain termination method. Next generation sequencing now allows millions of DNA molecules to be sequenced in parallel through massively parallel sequencing technologies.
DNA sequencing methods
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.
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.
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Dna sequencing
DNA sequencing determines the order of nucleotide bases in a DNA sample. The Sanger method, also known as dideoxy or chain termination method, was the first widely used DNA sequencing technique. It involves DNA polymerase, dNTPs, a primer, and ddNTPs to terminate DNA strand extension. The fragments are separated by gel electrophoresis and the sequence is read. Next-generation sequencing methods like pyrosequencing do not require electrophoresis or fragment separation. They detect incorporated nucleotides in real-time. Nanopore sequencing detects changes in electrical current as single-stranded DNA passes through a nanopore protein.
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An introduction to RNA-seq data analysis
AGRF in conjunction with EMBL Australia recently organised a workshop at Monash University Clayton. This workshop was targeted at beginners and biologists who are new to analysing Next-Gen Sequencing data. The workshop also aimed to provide users with a snapshot of bioinformatics and data analysis tips on how to begin to analyse project data. An introduction to RNA-seq data analysis was presented by AGRF Senior Bioinformatician Dr. Sonika Tyagi.
Presented: 1st August 2012
Dna sequencing
DNA sequencing is a process to determine the order of nucleotides in a DNA molecule. It was discovered in the 1970s by scientists like Frederick Sanger who developed the chain termination method. This method involves DNA replication with modified nucleotides that cause the growing DNA strand to terminate at that point. The fragments are then separated by size to reveal the sequence. Automated sequencing now uses fluorescent dyes and capillary electrophoresis for faster and higher throughput sequencing. DNA sequencing has applications in medicine, forensics, and agriculture.
Next generation sequencing
Next generation sequencing techniques have revolutionized DNA sequencing by increasing throughput and decreasing costs compared to previous methods like Sanger sequencing. Some key next generation sequencing methods include 454 sequencing (pyrosequencing), ABI Solid sequencing (sequencing by ligation), Illumina/Solexa sequencing (sequencing by synthesis), and nanopore sequencing. These new techniques allow for faster and cheaper large-scale sequencing and have enabled applications like whole genome sequencing.
Mapping the genome of bacteria
Genome mapping involves locating genes on chromosomes and determining the relative distances between genes. There are two main types of maps: genetic linkage maps which show the arrangement of genes based on their inheritance patterns, and physical maps which provide actual distances between landmarks on chromosomes. Physical maps can be further divided into cytogenetic maps, radiation hybrid maps, and sequence maps, with the complete DNA sequence being the ultimate physical map. Mapping methods include linkage analysis using genetic markers, as well as transformation, transduction, and sequencing of bacterial artificial chromosomes (BACs) and yeast artificial chromosomes (YACs).
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.
Microarray of long oligonucleotide
An oligonucleotide microarray consists of short DNA sequences attached to a solid surface to detect complementary DNA or RNA sequences in samples. Oligonucleotides are 13-25 nucleotide probes used to hybridize to specific gene sequences. In a microarray, fluorescently labeled sample sequences that bind to probes are detected by a scanner. This allows analysis of gene expression profiles or studies of gene function through comparative hybridization of normal versus experimental gene sequences.
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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.
Physical maps and their use in annotations
This document discusses physical maps and their use in genome annotation. It provides information on several key topics:
- Physical maps show the relative positions of genes on chromosomes, similar to a topological map of a country. They are created by identifying DNA fragments using genetic markers or restriction enzymes.
- Genetic mapping was first described in 1911 and applied to humans in the 1950s. Whole genome maps were generated by the mid-1990s using improved techniques.
- Physical mapping involves cloning chromosomal fragments, determining their sizes and relative locations to construct a map. Pulsed-field gel electrophoresis and fluorescence-activated cell sorting are used to isolate individual chromosomes.
- Contigs are assembled from overlapping cloned fragments to
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2. Total RNA is first extracted using Trizol, which separates RNA, DNA and proteins. mRNA is then isolated from total RNA using biotin-labeled oligo dT probes that selectively bind to the poly-A tail of mRNA molecules.
3. The mRNA-probe complexes are immobilized on magnetic beads coated with streptavidin. This allows the mRNA to be separated and purified from other RNAs through magnetic separation washes. The purified mRNA can then be used in applications like RT-PCR and protein
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Single nucleotide polymorphisms are single base variations between genomes within a species.
There are at least 10 million polymorphic sites in the human genome.
SNPs can distinguish individuals from one another
Denaturing Gradient Gel Electrophoresis
Chemical Cleavage Of Mismatch
Single-stranded Conformation Polymorphism (SSCP)
MutS Protein-binding Assays
Mismatch Repair Detection (MRD)
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Allele-specific oligonucleotide probes
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Targeting induced local lesions in genomes (TILLinG)
Minisequencing primers
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Pyrosequencing
This document discusses two approaches to pyrosequencing technology - solid phase and liquid phase. The solid phase approach utilizes streptavidin coated beads to immobilize biotin-labeled DNA templates. It involves sequential addition of nucleotides followed by washing steps to remove unincoporated nucleotides. The liquid phase approach introduced an enzyme called apyrase that degrades unincorporated nucleotides, eliminating the need for washing steps. It involves a cascade of four enzymes - DNA polymerase, ATP sulfurylase, luciferase, and apyrase to continuously degrade unincorporated nucleotides and determine DNA sequences from light signals.
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Maxam gilbert sequencing method
The Maxam-Gilbert sequencing method involves chemically cleaving DNA strands at specific bases, labeling the fragments radioactively, and separating them via gel electrophoresis. The sequence can then be read from the banding pattern on X-ray film. This early chemical degradation method was developed in 1977 and provided one of the first reliable ways to determine DNA sequences, but it uses toxic chemicals and radioactivity, has technical complexity, and can only read up to 500 base pairs.
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DNA Sequencing.pdf
DNA consists of a linear string of nucleotides, or bases, for simplicity, referred to by the first letters of their chemical names--A, T, C and G. The process of deducing the order of nucleotides in DNA is called DNA sequencing. Since the DNA sequence confers information that the cell uses to make RNA molecules and proteins, establishing the sequence of DNA is key for understanding how genomes work. The technology for DNA sequencing was made faster and less expensive as a part of the Human Genome Project. And recent developments have profoundly increased the efficiency of DNA sequencing even further.
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Next generation sequencing techniques have revolutionized DNA sequencing by increasing throughput and decreasing costs compared to previous methods like Sanger sequencing. Some key next generation sequencing methods include 454 sequencing (pyrosequencing), ABI Solid sequencing (sequencing by ligation), Illumina/Solexa sequencing (sequencing by synthesis), and nanopore sequencing. These new techniques allow for faster and cheaper large-scale sequencing and have enabled applications like whole genome sequencing.
Mapping the genome of bacteria
Genome mapping involves locating genes on chromosomes and determining the relative distances between genes. There are two main types of maps: genetic linkage maps which show the arrangement of genes based on their inheritance patterns, and physical maps which provide actual distances between landmarks on chromosomes. Physical maps can be further divided into cytogenetic maps, radiation hybrid maps, and sequence maps, with the complete DNA sequence being the ultimate physical map. Mapping methods include linkage analysis using genetic markers, as well as transformation, transduction, and sequencing of bacterial artificial chromosomes (BACs) and yeast artificial chromosomes (YACs).
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.
Microarray of long oligonucleotide
An oligonucleotide microarray consists of short DNA sequences attached to a solid surface to detect complementary DNA or RNA sequences in samples. Oligonucleotides are 13-25 nucleotide probes used to hybridize to specific gene sequences. In a microarray, fluorescently labeled sample sequences that bind to probes are detected by a scanner. This allows analysis of gene expression profiles or studies of gene function through comparative hybridization of normal versus experimental gene sequences.
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Ion Torrent (Proton/PGM) and SOLiD sequencing are two types of next-generation sequencing technologies. Ion Torrent uses semiconductor sequencing to detect hydrogen ions released during DNA synthesis, while SOLiD uses ligation of octamer probes and fluorescent dyes to determine sequences in color space. Both have advantages such as fast run times and high throughput but also limitations including errors in homopolymers for Ion Torrent and issues with palindromic sequences for SOLiD.
Dna sequencing
DNA sequencing is the process of determining the order of nucleotides in DNA. There are several methods of DNA sequencing including conventional, cycle sequencing, automated sequencing, and pyrosequencing. Conventional methods include chemical degradation and chain termination. Chemical degradation uses base-specific chemical reactions to cleave DNA fragments for sequencing. Chain termination uses DNA polymerase and dideoxynucleotides to terminate DNA strand extension for sequencing. Cycle sequencing applies the chain termination method to PCR for linear amplification of sequencing products. Automated sequencing uses fluorescence labeling for high-throughput sequencing. Pyrosequencing sequences DNA by detecting pyrophosphate release during polymerase nucleotide incorporation without electrophoresis.
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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.
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This document discusses physical maps and their use in genome annotation. It provides information on several key topics:
- Physical maps show the relative positions of genes on chromosomes, similar to a topological map of a country. They are created by identifying DNA fragments using genetic markers or restriction enzymes.
- Genetic mapping was first described in 1911 and applied to humans in the 1950s. Whole genome maps were generated by the mid-1990s using improved techniques.
- Physical mapping involves cloning chromosomal fragments, determining their sizes and relative locations to construct a map. Pulsed-field gel electrophoresis and fluorescence-activated cell sorting are used to isolate individual chromosomes.
- Contigs are assembled from overlapping cloned fragments to
mRNA Isolation
1. mRNA isolation is the process of extracting messenger RNA from biological samples. It is important for research and industry applications as mRNA provides insight into which genes are being expressed and translated into proteins.
2. Total RNA is first extracted using Trizol, which separates RNA, DNA and proteins. mRNA is then isolated from total RNA using biotin-labeled oligo dT probes that selectively bind to the poly-A tail of mRNA molecules.
3. The mRNA-probe complexes are immobilized on magnetic beads coated with streptavidin. This allows the mRNA to be separated and purified from other RNAs through magnetic separation washes. The purified mRNA can then be used in applications like RT-PCR and protein
SNP Detection Methods and applications
complete Single Nucleotide Polymorphiitsm Detection methods with Advance techniques with its applications
Single nucleotide polymorphisms are single base variations between genomes within a species.
There are at least 10 million polymorphic sites in the human genome.
SNPs can distinguish individuals from one another
Denaturing Gradient Gel Electrophoresis
Chemical Cleavage Of Mismatch
Single-stranded Conformation Polymorphism (SSCP)
MutS Protein-binding Assays
Mismatch Repair Detection (MRD)
Heteroduplex Analysis (HA)
Denaturing High Performance Liquid Chromatography (DHPLC)
UNG-Mediated T-Sequencing
RNA-Mediated Finger printing with MALDI MS Detection
Sequencing by Hybridization
Direct DNA Sequencing
Single-feature polymorphism (SFP)
Invader probe
Allele-specific oligonucleotide probes
PCR-based methods
Allele specific primers
Sequence Polymorphism-Derived (SPD) markers
Targeting induced local lesions in genomes (TILLinG)
Minisequencing primers
Allele-specific ligation probes
Different pcr techniques and their application
This document discusses several types of PCR techniques and their applications. It begins by explaining standard PCR and its development. It then describes several specialized PCR techniques including allele-specific PCR, asymmetric PCR, assembly PCR, hot-start PCR, helicase-dependent amplification, in situ PCR, inverse PCR, ligation-mediated PCR, and multiplex ligation-dependent probe amplification. Each technique is explained and examples of its uses and applications are provided.
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This document discusses different methods for ligating DNA fragments with vectors, including DNA ligase, homopolymer tailing, linkers, and adaptors. It provides details on how each method works, such as using DNA ligase to join complementary cohesive ends, adding homopolymeric tails to fragments using terminal deoxynucleotidyl transferase, and how linkers and adaptors can be used to generate sticky ends for ligation. The basic difference between linkers and adaptors is that linkers have blunt ends while adaptors have cohesive ends.
Pyrosequencing
This document discusses two approaches to pyrosequencing technology - solid phase and liquid phase. The solid phase approach utilizes streptavidin coated beads to immobilize biotin-labeled DNA templates. It involves sequential addition of nucleotides followed by washing steps to remove unincoporated nucleotides. The liquid phase approach introduced an enzyme called apyrase that degrades unincorporated nucleotides, eliminating the need for washing steps. It involves a cascade of four enzymes - DNA polymerase, ATP sulfurylase, luciferase, and apyrase to continuously degrade unincorporated nucleotides and determine DNA sequences from light signals.
Dna fingerprinting
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Maxam gilbert sequencing method
The Maxam-Gilbert sequencing method involves chemically cleaving DNA strands at specific bases, labeling the fragments radioactively, and separating them via gel electrophoresis. The sequence can then be read from the banding pattern on X-ray film. This early chemical degradation method was developed in 1977 and provided one of the first reliable ways to determine DNA sequences, but it uses toxic chemicals and radioactivity, has technical complexity, and can only read up to 500 base pairs.
GENE ISOLATION AND SEQUENCING.pdf
DNA sequencing is the process of determining the nucleic acid sequence – the order of nucleotides in DNA. It includes any method or technology that is used to determine the order of the four bases: adenine, guanine, cytosine, and thymine.
Random Amplified Polymorphic Dna
This document summarizes the random amplified polymorphic DNA (RAPD) technique. RAPD is a type of PCR that uses random nucleotide primers to amplify unknown regions of genomic DNA. It was developed in 1991 and involves using single, short random primers to amplify random DNA segments. The amplified products are then separated via gel electrophoresis and visualized. RAPD is a quick and inexpensive molecular marker technique that requires no prior DNA sequence knowledge, but lacks reproducibility and produces dominant markers. Its applications include assessing genetic diversity, mapping genomes, and use in breeding and evolutionary studies.
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DNA modifying enzymes include nucleases, ligases, polymerases, phosphatases and other enzymes that modify DNA structure and sequence. Nucleases such as exonucleases and endonucleases cleave phosphodiester bonds in nucleic acids. Ligases join nucleic acid molecules by forming phosphodiester bonds. Polymerases such as DNA polymerase synthesize new DNA strands using existing strands as templates. These enzymes are essential for processes like DNA replication, repair and recombination.
DNA Sequencing.pdf
DNA consists of a linear string of nucleotides, or bases, for simplicity, referred to by the first letters of their chemical names--A, T, C and G. The process of deducing the order of nucleotides in DNA is called DNA sequencing. Since the DNA sequence confers information that the cell uses to make RNA molecules and proteins, establishing the sequence of DNA is key for understanding how genomes work. The technology for DNA sequencing was made faster and less expensive as a part of the Human Genome Project. And recent developments have profoundly increased the efficiency of DNA sequencing even further.
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Notes how to work with variables, constants and do calculations
Notes how to work with variables, constants and do calculations
Similar to Dna sequencing
BIOTECHNOLOGY PPT.pptx
Genomics is the study of genomes through DNA sequencing and analysis. There are several types of genomics including structural, functional, comparative, epigenomics, and metagenomics. DNA sequencing methods have advanced from Sanger sequencing to next-generation sequencing using high-throughput technologies. Genome sequencing has provided insights into disease, evolution, and applications in medicine, agriculture, and environmental science.
theoretical perspectives on marriage and family
This document provides an overview of DNA sequencing, including its definition, types, uses, and impact. It discusses the key types of sequencing, including the Sanger method and high-throughput sequencing techniques like Illumina and Roche 454. The document also outlines some common uses of sequencing in areas like diagnostics, genomics, and forensics. Overall, the document highlights how advances in sequencing technology have enabled sequencing of entire genomes and provided important insights into human biology, disease, and more.
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
Different methods of gene sequencing durgesh sirohi
The document discusses various methods of gene sequencing. It begins with classical methods like Maxam-Gilbert chemical degradation and Sanger dideoxy chain termination. It then covers advanced methods such as shotgun sequencing and bridge PCR. The document also describes several new generation sequencing techniques, including sequencing by synthesis, pyrosequencing, nanopore sequencing, ion torrent sequencing, and Illumina sequencing. It provides details on the principles and procedures of many of these sequencing methods.
Gene Sequencing | maxam gilbert sequencing | sanger sequencing
NEED OF GENETIC SEQUENCING
- Understanding the particular DNA sequence can shed light on a genetic condition and offer hope for the eventual development of treatment.
- An alteration in a DNA sequence can lead to an altered or non functional protein and hence to a harmful effect in a plant or animal.
- Simple point mutations can cause altered protein shape and function.
-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
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.
Describe the steps in Sanger DNA sequencing. Provide a high level des.pdf
Consider the program: var s: int:= 1, i: int co i:= 1 to 2 rightarrow do true rightarrow (await s >
0 rightarrow s: = s - 1) (s:= s + 1) Above, S_i is a statement list that is assumed not to modify
shared variable s. Develop complete proof outlines for the two processes. Demonstrate that the
proofs of the processes are interference-free. Then use the proof outlines and the method of
Exclusion of Configurations (2.25) to show that S_1 and S_2 cannot execute at the same time
and that the program is deadlock-free. What scheduling policy is required to ensure that a
process delayed at its first await statement will eventually be able to proceed? Explain.
Solution
int i=1
do(i=1 to 2)
{
wait(s>0){
s=s-1
Si
}
s=s+1
}
to prove that it should be deadlock free it should support mutual exclusion,progress,bounded
waiting.
mutual exclusion:-
p1:
int i=1
do(i=1 to 2) //entry section
{
wait(s>0){ //critical section
s=s-1
Si
}
s=s+1
} //exit section
so while p1 is exexcuting p2 while s enters into critical section as we don\'t update any values so
mutual exclusion is possible and can be formed.
progress:-
when p1 is in non-critical section p2can execute in critical section.program gurantees progress.
bounded-waiting :-
suitable for ony countable no of procedures and processes.
as it satisfies all properties ....so it\'s deadlock free..
DNA MUGSHOTS AGAINST CRIME
The document discusses DNA and its use in species identification and forensics. It notes there are estimated to be 8.7 million species on Earth. DNA contains the genetic code and instructions for making proteins. DNA profiling uses restriction enzymes to cut DNA into fragments that can then be compared to identify individuals. Advances allow DNA to be extracted from smaller samples and new sources. DNA evidence has limitations but continues to be a powerful forensic tool when handled properly.
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.
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.
DNA sequencing
This document discusses various methods for DNA sequencing, including both early chemical methods and more recent next-generation sequencing technologies. It describes Frederick Sanger's chain termination method from 1977, which uses DNA polymerase, dNTPs, and ddNTPs. It also describes Maxam-Gilbert sequencing from the same year, which uses chemical cleavage. Next-generation methods discussed include pyrosequencing, sequencing by ligation, sequencing by synthesis, and ion semiconductor sequencing. The document compares the different methods based on read length, accuracy, throughput, run time, and cost.
Lecture 3 southern blotting
This document discusses blotting techniques, specifically Southern blotting. It defines Southern blotting as a technique developed by Professor Sir Edwin Southern in 1975 to detect specific DNA sequences. The technique involves separating DNA fragments via gel electrophoresis, transferring them to a membrane, then using a labeled probe to detect complementary DNA sequences via hybridization. Southern blotting is used for applications like gene discovery, mapping, diagnostics, and DNA fingerprinting.
DNA SEQUENCING (1).pptx
DNA sequencing is the process of determining the nucleic acid sequence – the order of nucleotides in DNA. It includes any method or technology that is used to determine the order of the four bases: adenine, guanine, cytosine, and thymine. The advent of rapid DNA sequencing methods has greatly accelerated biological and medical research and discovery.
Knowledge of DNA sequences has become indispensable for basic biological research, DNA Genographic Projects and in numerous applied fields such as medical diagnosis, biotechnology, forensic biology, virology and biological systematics. Comparing healthy and mutated DNA sequences can diagnose different diseases including various cancers,characterize antibody repertoire, and can be used to guide patient treatment.[5Having a quick way to sequence DNA allows for faster and more individualized medical care to be administered, and for more organisms to be identified and cataloged.
The rapid speed of sequencing attained with modern DNA sequencing technology has been instrumental in the sequencing of complete DNA sequences, or genomes, of numerous types and species of life, including the human genome and other complete DNA sequences of many animal, plant, and microbial species.
The first DNA sequences were obtained in the early 1970s by academic researchers using laborious methods based on two-dimensional chromatography. Following the development of fluorescence-based sequencing methods with a DNA sequencer, DNA sequencing has become easier and orders of magnitude faster.
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.Whole Genome Sequencing
•Allows doctors to closely analyze a patient's genes for mutations and health indicators.
•Can detect intellectual disabilities and developmental delays.
•WGS is currently available at Yale for patients in the NICU and PICU.
•Involves Genetics.Sequencing may be utilized to determine the order of nucleotides in small targeted genomic regions or entire genomes. Illumina sequencing enables a wide variety of applications, allowing researchers to ask virtually any question related to the genome, transcriptome, or epigenome of any organism.The spectrum of analysis of NGS can extend from a small number of genes to an entire genome, depending on the goal. Whole-genome sequencing (WGS) and whole-exome sequencing (WES) provide the sequence of DNA bases across the genome and exome, respectively.Capillary electrophoresis (CE) instruments are capable of performing both Sanger sequencing and fragment analysis. Fragment analysis is a method in which DNA fragments are fluorescently labeled, separated by CE, and sized by comparison to an internal standard. sanger and Maxam-Gilbert sequencing technologies were classified
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 Fingerprinting
DNA fingerprinting is a technique used for identification by extracting and analyzing the base pair pattern of an individual's DNA. It involves isolating DNA from a sample, cutting the DNA into fragments using restriction enzymes, and comparing the fragment patterns on a gel to identify individuals. The main applications of DNA fingerprinting are in solving criminal cases by matching DNA evidence to suspects, diagnosing inherited diseases, and determining biological relationships in areas like paternity testing.
Gen sequencing strategies by kk sahu
INTRODUCTION
HISTORY
WHY GENOME SEQUENCE IS IMPORTANT
METHODS FOR SEQUENCING
MAXAM & GILBERT’S METHOD
SANGER’S METHOD
AUTOMATIC SEQUENCING
SHOTGUN SEQUENCING
APPLICATION
CONCLUSION
REFERENCE
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.
Molecular hybridization
Molecular hybridization is the process by which two complementary strands of DNA or RNA bind together via hydrogen bonding between bases. It is used in techniques like cloning, PCR, and diagnostic tests involving nucleic acid probes. The document describes the process of hybridization, factors that affect binding strength, and techniques that utilize molecular hybridization like Southern blotting, dot/slot blotting, microarrays, and in situ hybridization.
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Different methods of gene sequencing durgesh sirohi
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Dna sequencing
- 1. Neeraj Sharma MSc. Medical Microbiology 2nd year
- 2. The process of determining the order of the nucleotide bases along a DNA strand. In 1977 two separate methods of sequencing of DNA were developed. Chain termination method and Chemical Degredation method. Chain termination method is commonly used today
- 3. Also called dideoxynucleotide or Sangers method. This method is based on the principle that single stranded DNA molecules that differ in length by just a nucleotide can be separated from one another using polyacrylamide gel electrophoresis.
- 4. A DNA template. A primer tagged with a mildly radioactive molecule or light emitting chemical. DNA Polymerase Deoxynucleotide (G, A, C, T) Dideoxynucleotide (ddG, ddA, ddC, ddT)
- 5. Steps: 1. Denaturation 2. Primer attachment and extension of bases 3. Termination 4. Gel electrophoresis
- 6. Denaturation of the Double stranded DNA into a single stranded DNA template.
- 7. Carried out in four set of test tubes. Each tube contains DNA fragments, DNA primer, nucleotides, DNA polymerase. 1% of the total solution ddA, ddG, ddC and ddT in separate tubes.
- 10. The DNA fragments are separated by size with polyacrylamide gel electrophoresis , and because a radio active DNA primer was used , the position of the band in the gel is determined by autoradiography.
- 11. The autoradiogram is then read starting at the bottom where the DNA fragments are the shortest. These bands at the bottom represent the sequence of DNA bases nearest to the primer.
- 12. The sequence is read one base at a time in the 5’ to 3’ direction. The sequence being read is complementary to the DNA fragment. The sequence of short fragment is 5’ A G C C T A G A C T 3’
- 13. Other methods. Shot gun sequencing. Pyrosequencing.
- 14. Determine the sequence of individual gene, larger genetic regions, full chromosome or entire genome. Provide the order of individual nucleotides in DNA RNA isolated from cells of animals plants bacteria etc.
- 15. Molecular biology - studying the genome itself, how proteins are made, what proteins are made, identifying new genes and associations with diseases and phenotypes, and identifying potential drug targets Evolutionary biology - studying how different organisms are related and how they evolved Helpful in ecology, epidemiology,research and other fields.