This document provides an introduction to DNA fingerprinting and forensic analysis. It discusses how DNA fragments show unique patterns that can be used for paternity disputes and forensic evidence. The document describes the process of preparing a DNA fingerprint, including specimen collection and storage, as well as restriction fragment length polymorphism (RFLP) and variable number tandem repeats (VNTR) analysis. It also explains techniques like the Southern blot, polymerase chain reaction (PCR), and how DNA fingerprinting can be applied to diagnosing disease, paternity testing, forensics, and genealogy.
This document discusses DNA fingerprinting and forensic analysis techniques. It describes how DNA fragments show unique patterns that can be used for paternity disputes and forensic evidence. The key techniques discussed are restriction fragment length polymorphism (RFLP) which separates DNA fragments by size, Southern blotting which transfers DNA fragments to a membrane for probing, variable number tandem repeats (VNTR) which are repeated DNA sequences that vary between individuals, and polymerase chain reaction (PCR) which amplifies specific DNA regions. These DNA analysis techniques have various applications including diagnosing disease, paternity testing, forensics, and genealogy.
Sanger sequencing is one of the DNA sequencing methods used to identify and determine the sequence (Nucleotide) of DNA .This is an enzymatic method of sequencing developed by Fred Sanger.
The methods used for DNA finger printing are the same Molecular markers...so for detailed note on the steps which is explained in DNA typing can be used to study the performance pf markers too...
DNA fingerprinting is a technique developed in 1984 that analyzes variable regions in genetic material to distinguish one person from another. It involves isolating DNA from a sample, cutting it with restriction enzymes, sorting by size, and probing specific locations to generate a unique pattern. This technique uses variations in the number of short tandem repeats between individuals and has been used successfully in criminal cases and establishing paternity. The most common DNA fingerprinting methods are electrophoresis, polymerase chain reaction, restriction fragment length polymorphism, random amplified polymorphic DNA, and amplified fragment length polymorphism. DNA fingerprinting has applications in diagnosing inherited disorders, developing cures, and identifying criminals using biological evidence.
DNA profiling uses patterns of repeating DNA sequences that vary between individuals to identify people at a molecular level. It has various applications including criminal identification by comparing DNA evidence from a crime scene to a suspect's DNA, determining biological relationships in cases of disputed parentage, and diagnosing inherited genetic disorders. The most common current method is short tandem repeat analysis using PCR to amplify short repeating sequences and generate a DNA fingerprint.
Polymerase chain reaction (PCR) is a method to rapidly amplify a specific DNA sample, allowing scientists to study very small amounts of DNA. PCR uses the enzyme DNA polymerase to assemble copies of DNA by denaturing and separating the DNA strands, annealing primers to the strands, and extending the primers to make new copies of DNA. This cycling process can produce millions of copies of DNA. PCR is used in applications like disease diagnosis, forensics, genetic testing, and evolutionary studies.
This document summarizes DNA fingerprinting, which involves analyzing variable tandem repeat regions in DNA to generate unique genetic profiles for identification purposes. It describes how DNA is extracted from samples, cut using restriction enzymes, separated via gel electrophoresis, and analyzed using probes to develop a fingerprint. Key applications include identifying suspects in criminal cases, solving paternity disputes, diagnosing genetic disorders, and personal identification. The technique was pioneered in 1984 by Alec Jeffreys and first used in court in 1987. Famous cases that have utilized DNA fingerprinting include identifying the killer in the Colin Pitchfork murder case and establishing Steve Bing's paternity of Elizabeth Hurley's son.
This document discusses molecular genetic methods such as polymerase chain reaction (PCR), DNA sequencing, DNA fingerprinting, and single nucleotide polymorphisms. It provides details on how each method works, including how PCR amplifies DNA, the process of manual and automated DNA sequencing, using variable number tandem repeats as markers for DNA fingerprinting, and applications of these molecular genetic techniques.
This document discusses DNA fingerprinting and forensic analysis techniques. It describes how DNA fragments show unique patterns that can be used for paternity disputes and forensic evidence. The key techniques discussed are restriction fragment length polymorphism (RFLP) which separates DNA fragments by size, Southern blotting which transfers DNA fragments to a membrane for probing, variable number tandem repeats (VNTR) which are repeated DNA sequences that vary between individuals, and polymerase chain reaction (PCR) which amplifies specific DNA regions. These DNA analysis techniques have various applications including diagnosing disease, paternity testing, forensics, and genealogy.
Sanger sequencing is one of the DNA sequencing methods used to identify and determine the sequence (Nucleotide) of DNA .This is an enzymatic method of sequencing developed by Fred Sanger.
The methods used for DNA finger printing are the same Molecular markers...so for detailed note on the steps which is explained in DNA typing can be used to study the performance pf markers too...
DNA fingerprinting is a technique developed in 1984 that analyzes variable regions in genetic material to distinguish one person from another. It involves isolating DNA from a sample, cutting it with restriction enzymes, sorting by size, and probing specific locations to generate a unique pattern. This technique uses variations in the number of short tandem repeats between individuals and has been used successfully in criminal cases and establishing paternity. The most common DNA fingerprinting methods are electrophoresis, polymerase chain reaction, restriction fragment length polymorphism, random amplified polymorphic DNA, and amplified fragment length polymorphism. DNA fingerprinting has applications in diagnosing inherited disorders, developing cures, and identifying criminals using biological evidence.
DNA profiling uses patterns of repeating DNA sequences that vary between individuals to identify people at a molecular level. It has various applications including criminal identification by comparing DNA evidence from a crime scene to a suspect's DNA, determining biological relationships in cases of disputed parentage, and diagnosing inherited genetic disorders. The most common current method is short tandem repeat analysis using PCR to amplify short repeating sequences and generate a DNA fingerprint.
Polymerase chain reaction (PCR) is a method to rapidly amplify a specific DNA sample, allowing scientists to study very small amounts of DNA. PCR uses the enzyme DNA polymerase to assemble copies of DNA by denaturing and separating the DNA strands, annealing primers to the strands, and extending the primers to make new copies of DNA. This cycling process can produce millions of copies of DNA. PCR is used in applications like disease diagnosis, forensics, genetic testing, and evolutionary studies.
This document summarizes DNA fingerprinting, which involves analyzing variable tandem repeat regions in DNA to generate unique genetic profiles for identification purposes. It describes how DNA is extracted from samples, cut using restriction enzymes, separated via gel electrophoresis, and analyzed using probes to develop a fingerprint. Key applications include identifying suspects in criminal cases, solving paternity disputes, diagnosing genetic disorders, and personal identification. The technique was pioneered in 1984 by Alec Jeffreys and first used in court in 1987. Famous cases that have utilized DNA fingerprinting include identifying the killer in the Colin Pitchfork murder case and establishing Steve Bing's paternity of Elizabeth Hurley's son.
This document discusses molecular genetic methods such as polymerase chain reaction (PCR), DNA sequencing, DNA fingerprinting, and single nucleotide polymorphisms. It provides details on how each method works, including how PCR amplifies DNA, the process of manual and automated DNA sequencing, using variable number tandem repeats as markers for DNA fingerprinting, and applications of these molecular genetic techniques.
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.
The document summarizes molecular diagnostic techniques for detecting plant pathogens. It discusses several techniques including polymerase chain reaction (PCR), molecular hybridization, molecular markers, nucleic acid sequencing, and microarrays. PCR is described as a sensitive technique that can amplify DNA from pathogens. Molecular hybridization uses probes to detect complementary DNA or RNA sequences from pathogens. Molecular markers like RFLP and RAPD can identify pathogens by detecting DNA polymorphisms. Nucleic acid sequencing techniques like NASBA and LAMP can detect and amplify RNA from pathogens. Microarrays allow simultaneous detection of multiple pathogens using DNA probes spotted onto a chip.
DNA fingerprinting is a technique that analyzes variations in DNA sequences at specific locations in the genome to identify individuals. There are two main methods: RFLP (restriction fragment length polymorphism) and PCR (polymerase chain reaction). RFLP involves digesting DNA with restriction enzymes, separating fragments by size, and detecting with probes. PCR amplifies specific DNA regions defined by primer sequences. Short tandem repeats (STRs) are now commonly analyzed by PCR. DNA fingerprinting is used in criminal investigations to identify suspects or victims, and in resolving medical issues like paternity disputes. DNA databases help law enforcement match crime scene evidence to suspects.
The document discusses polymerase chain reaction (PCR), a laboratory technique used to amplify a specific segment of DNA. It was invented in 1983 by Kary Mullis, who was awarded the Nobel Prize in 1993. PCR works by repeating cycles of heating and cooling of the DNA sample to separate the double helix, followed by use of DNA polymerase to make copies of the targeted region. This process can generate millions of copies of the targeted DNA sequence. The document outlines the components and steps of PCR, including primers, DNA polymerase, and thermal cycling. It discusses some applications of PCR such as detecting low-abundance DNA sequences, forensic analysis, and prenatal diagnosis.
Polymerase Chain Reaction (PCR) is an in vitro technique for amplifying specific DNA sequences. It involves repeated cycles of denaturation, annealing of primers to the DNA templates, and extension of the primers by DNA polymerase. This allows for exponential amplification of the target DNA region. PCR has many applications in medicine, forensics, and research, such as detecting genetic diseases or infectious agents, genetic fingerprinting, and studying gene expression and genomes. It is a powerful technique that can generate millions of copies of DNA from a single template.
The document discusses the polymerase chain reaction (PCR) technique. It describes how PCR allows targeted amplification of specific DNA regions. Key steps in PCR include using DNA polymerase, primers, and thermal cycling to denature and replicate the DNA. The document outlines different types of PCR like quantitative, multiplex, and nested PCR. It also discusses how PCR has revolutionized fields like molecular archaeology by enabling the cloning of ancient DNA. PCR is now widely used in applications like forensics, disease diagnosis, and genome sequencing.
This document discusses DNA fingerprinting techniques including restriction fragment length polymorphism (RFLP) and DNA footprinting. RFLP involves using restriction enzymes to cut DNA at specific sites, resulting in fragments of varying lengths that can be used to differentiate individuals. DNA footprinting identifies the specific binding sites of DNA-binding proteins by detecting regions of DNA that are protected from cleavage by bound proteins. The document provides detailed explanations of the principles and procedures of these techniques.
This document provides an overview of DNA structure and function. It discusses how DNA is organized into chromosomes and genes within cells. The key components of DNA, including nucleotides, the sugar-phosphate backbone, and the double helix structure are described. The functions of DNA in transmitting genetic information from generation to generation and providing blueprints for protein synthesis are highlighted. Methods for analyzing DNA like restriction enzymes, gel electrophoresis, PCR, and DNA fingerprinting are summarized. Uses of DNA analysis in forensic investigations and criminal databases are also mentioned.
What is PCR?
History of PCR
Components of PCR
Principles of PCR
Basic Requirements
Instrumentation
PCR Programme
Advantages of PCR
Applications of PCR
Conclusion
References
The document discusses various molecular cytogenetics techniques including polymerase chain reaction (PCR), reverse transcription PCR (RT-PCR), and fluorescent in situ hybridization (FISH). It provides details on the principles, techniques, and applications of PCR and RT-PCR. PCR is described as a technique that amplifies specific DNA regions, allowing minute quantities to be analyzed. Key steps involve DNA denaturation, primer annealing, and fragment extension. RT-PCR involves first converting RNA to cDNA then amplifying a specific region. Both techniques have numerous diagnostic and research applications.
The polymerase chain reaction (PCR) is a technique used to quickly make millions of copies of a specific DNA sequence. The process involves:
1. Denaturing the DNA template into single strands by heating.
2. Adding primers that are complementary to the DNA sequences and allowing them to anneal.
3. Using DNA polymerase to synthesize new DNA strands.
4. Repeating the process many times to exponentially amplify the target DNA sequence.
PCR has revolutionized DNA analysis and is used for applications like genetic testing, fingerprinting, cloning, and DNA sequencing. It relies on thermostable DNA polymerases that can function at high temperatures needed for denaturing.
This document provides an overview of polymerase chain reaction (PCR). It discusses:
1. The key discoveries and scientists that developed PCR, including Kary Mullis who won the 1993 Nobel Prize for his conception of PCR.
2. The basic steps and components of PCR including DNA template, primers, Taq polymerase enzyme, repeated temperature cycling of denaturation, annealing and extension to exponentially amplify the target DNA region.
3. Applications of PCR including DNA cloning, medical diagnostics, forensics, and research. PCR is used to amplify specific genes or DNA regions for various downstream analyses and applications.
Genetic engineering involves manipulating genetic material (DNA) to achieve desired goals. The basic principles involve artificially copying DNA from one organism and joining it into the DNA of another. Molecular tools like restriction enzymes and DNA ligases are used to cut and join DNA. Methods to transfer genes include transformation, electroporation, and liposome-mediated transfer. Applications include producing human proteins like insulin, developing gene therapies, and genetically modifying plants. Gene libraries, blotting techniques like Southern blotting, and PCR are also discussed as important molecular tools in genetic engineering.
DNA profiling is a technique used to distinguish individuals using DNA samples. It was invented in 1985 by Alec Jeffreys. The process involves breaking down cells to extract DNA, cutting the DNA into fragments using restriction enzymes, separating the fragments by size using gel electrophoresis, and analyzing the pattern of fragments to obtain a DNA profile. DNA profiling is used in criminal investigations to identify suspects by comparing crime scene DNA to suspects' profiles. It is also used to determine biological relationships in cases of paternity, maternity, and inheritance disputes. Famous cases that utilized DNA profiling include identifying Colin Pitchfork as the first criminal caught this way and the O.J. Simpson murder trial.
PCR is a technique used to amplify a specific DNA sequence. It involves repeated cycles of heating and cooling of the DNA sample to denature and separate the DNA strands, followed by primer annealing and polymerase extension. This results in exponential amplification of the target DNA sequence. PCR requires a DNA template, DNA polymerase, primers, nucleotides, and repeated cycling between high and low temperatures. It has applications in research, forensics, medicine and molecular biology.
To describe DNA extraction
To explain and demonstrate DNA cloning
To explain the process of PCR and its uses.
To explain DNA fingerprinting and its uses
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.
PCR is a technique used to amplify a specific region of DNA. It involves three main steps - denaturation to separate the DNA strands, annealing where primers attach to the DNA, and elongation where a DNA polymerase enzyme copies the DNA. During each cycle of PCR, the amount of target DNA doubles, allowing millions of copies to be produced from a single DNA molecule. PCR is a powerful tool used in various fields like forensics, medicine, and research.
The document summarizes the history and development of PCR technology. It describes key events like the concept of PCR being presented in 1984, the first PCR device called Mr. Cycle in 1985, the isolation of Taq polymerase in 1985 which enabled PCR without manual addition of reagents, and the licensing of PCR by Roche in 1991 for commercial use. It also provides details on the basic components and process of PCR.
Southern blotting and Northern blotting are techniques used to detect specific DNA and RNA sequences. Southern blotting involves transferring DNA fragments separated by gel electrophoresis to a membrane, then using a labeled probe to identify fragments by hybridization. Northern blotting follows similar steps but detects RNA, requiring formaldehyde treatment to denature RNA. Both techniques allow identification of specific molecules in complex mixtures.
Exploiting Artificial Intelligence for Empowering Researchers and Faculty, In...Dr. Vinod Kumar Kanvaria
Exploiting Artificial Intelligence for Empowering Researchers and Faculty,
International FDP on Fundamentals of Research in Social Sciences
at Integral University, Lucknow, 06.06.2024
By Dr. Vinod Kumar Kanvaria
Assessment and Planning in Educational technology.pptxKavitha Krishnan
In an education system, it is understood that assessment is only for the students, but on the other hand, the Assessment of teachers is also an important aspect of the education system that ensures teachers are providing high-quality instruction to students. The assessment process can be used to provide feedback and support for professional development, to inform decisions about teacher retention or promotion, or to evaluate teacher effectiveness for accountability purposes.
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.
The document summarizes molecular diagnostic techniques for detecting plant pathogens. It discusses several techniques including polymerase chain reaction (PCR), molecular hybridization, molecular markers, nucleic acid sequencing, and microarrays. PCR is described as a sensitive technique that can amplify DNA from pathogens. Molecular hybridization uses probes to detect complementary DNA or RNA sequences from pathogens. Molecular markers like RFLP and RAPD can identify pathogens by detecting DNA polymorphisms. Nucleic acid sequencing techniques like NASBA and LAMP can detect and amplify RNA from pathogens. Microarrays allow simultaneous detection of multiple pathogens using DNA probes spotted onto a chip.
DNA fingerprinting is a technique that analyzes variations in DNA sequences at specific locations in the genome to identify individuals. There are two main methods: RFLP (restriction fragment length polymorphism) and PCR (polymerase chain reaction). RFLP involves digesting DNA with restriction enzymes, separating fragments by size, and detecting with probes. PCR amplifies specific DNA regions defined by primer sequences. Short tandem repeats (STRs) are now commonly analyzed by PCR. DNA fingerprinting is used in criminal investigations to identify suspects or victims, and in resolving medical issues like paternity disputes. DNA databases help law enforcement match crime scene evidence to suspects.
The document discusses polymerase chain reaction (PCR), a laboratory technique used to amplify a specific segment of DNA. It was invented in 1983 by Kary Mullis, who was awarded the Nobel Prize in 1993. PCR works by repeating cycles of heating and cooling of the DNA sample to separate the double helix, followed by use of DNA polymerase to make copies of the targeted region. This process can generate millions of copies of the targeted DNA sequence. The document outlines the components and steps of PCR, including primers, DNA polymerase, and thermal cycling. It discusses some applications of PCR such as detecting low-abundance DNA sequences, forensic analysis, and prenatal diagnosis.
Polymerase Chain Reaction (PCR) is an in vitro technique for amplifying specific DNA sequences. It involves repeated cycles of denaturation, annealing of primers to the DNA templates, and extension of the primers by DNA polymerase. This allows for exponential amplification of the target DNA region. PCR has many applications in medicine, forensics, and research, such as detecting genetic diseases or infectious agents, genetic fingerprinting, and studying gene expression and genomes. It is a powerful technique that can generate millions of copies of DNA from a single template.
The document discusses the polymerase chain reaction (PCR) technique. It describes how PCR allows targeted amplification of specific DNA regions. Key steps in PCR include using DNA polymerase, primers, and thermal cycling to denature and replicate the DNA. The document outlines different types of PCR like quantitative, multiplex, and nested PCR. It also discusses how PCR has revolutionized fields like molecular archaeology by enabling the cloning of ancient DNA. PCR is now widely used in applications like forensics, disease diagnosis, and genome sequencing.
This document discusses DNA fingerprinting techniques including restriction fragment length polymorphism (RFLP) and DNA footprinting. RFLP involves using restriction enzymes to cut DNA at specific sites, resulting in fragments of varying lengths that can be used to differentiate individuals. DNA footprinting identifies the specific binding sites of DNA-binding proteins by detecting regions of DNA that are protected from cleavage by bound proteins. The document provides detailed explanations of the principles and procedures of these techniques.
This document provides an overview of DNA structure and function. It discusses how DNA is organized into chromosomes and genes within cells. The key components of DNA, including nucleotides, the sugar-phosphate backbone, and the double helix structure are described. The functions of DNA in transmitting genetic information from generation to generation and providing blueprints for protein synthesis are highlighted. Methods for analyzing DNA like restriction enzymes, gel electrophoresis, PCR, and DNA fingerprinting are summarized. Uses of DNA analysis in forensic investigations and criminal databases are also mentioned.
What is PCR?
History of PCR
Components of PCR
Principles of PCR
Basic Requirements
Instrumentation
PCR Programme
Advantages of PCR
Applications of PCR
Conclusion
References
The document discusses various molecular cytogenetics techniques including polymerase chain reaction (PCR), reverse transcription PCR (RT-PCR), and fluorescent in situ hybridization (FISH). It provides details on the principles, techniques, and applications of PCR and RT-PCR. PCR is described as a technique that amplifies specific DNA regions, allowing minute quantities to be analyzed. Key steps involve DNA denaturation, primer annealing, and fragment extension. RT-PCR involves first converting RNA to cDNA then amplifying a specific region. Both techniques have numerous diagnostic and research applications.
The polymerase chain reaction (PCR) is a technique used to quickly make millions of copies of a specific DNA sequence. The process involves:
1. Denaturing the DNA template into single strands by heating.
2. Adding primers that are complementary to the DNA sequences and allowing them to anneal.
3. Using DNA polymerase to synthesize new DNA strands.
4. Repeating the process many times to exponentially amplify the target DNA sequence.
PCR has revolutionized DNA analysis and is used for applications like genetic testing, fingerprinting, cloning, and DNA sequencing. It relies on thermostable DNA polymerases that can function at high temperatures needed for denaturing.
This document provides an overview of polymerase chain reaction (PCR). It discusses:
1. The key discoveries and scientists that developed PCR, including Kary Mullis who won the 1993 Nobel Prize for his conception of PCR.
2. The basic steps and components of PCR including DNA template, primers, Taq polymerase enzyme, repeated temperature cycling of denaturation, annealing and extension to exponentially amplify the target DNA region.
3. Applications of PCR including DNA cloning, medical diagnostics, forensics, and research. PCR is used to amplify specific genes or DNA regions for various downstream analyses and applications.
Genetic engineering involves manipulating genetic material (DNA) to achieve desired goals. The basic principles involve artificially copying DNA from one organism and joining it into the DNA of another. Molecular tools like restriction enzymes and DNA ligases are used to cut and join DNA. Methods to transfer genes include transformation, electroporation, and liposome-mediated transfer. Applications include producing human proteins like insulin, developing gene therapies, and genetically modifying plants. Gene libraries, blotting techniques like Southern blotting, and PCR are also discussed as important molecular tools in genetic engineering.
DNA profiling is a technique used to distinguish individuals using DNA samples. It was invented in 1985 by Alec Jeffreys. The process involves breaking down cells to extract DNA, cutting the DNA into fragments using restriction enzymes, separating the fragments by size using gel electrophoresis, and analyzing the pattern of fragments to obtain a DNA profile. DNA profiling is used in criminal investigations to identify suspects by comparing crime scene DNA to suspects' profiles. It is also used to determine biological relationships in cases of paternity, maternity, and inheritance disputes. Famous cases that utilized DNA profiling include identifying Colin Pitchfork as the first criminal caught this way and the O.J. Simpson murder trial.
PCR is a technique used to amplify a specific DNA sequence. It involves repeated cycles of heating and cooling of the DNA sample to denature and separate the DNA strands, followed by primer annealing and polymerase extension. This results in exponential amplification of the target DNA sequence. PCR requires a DNA template, DNA polymerase, primers, nucleotides, and repeated cycling between high and low temperatures. It has applications in research, forensics, medicine and molecular biology.
To describe DNA extraction
To explain and demonstrate DNA cloning
To explain the process of PCR and its uses.
To explain DNA fingerprinting and its uses
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.
PCR is a technique used to amplify a specific region of DNA. It involves three main steps - denaturation to separate the DNA strands, annealing where primers attach to the DNA, and elongation where a DNA polymerase enzyme copies the DNA. During each cycle of PCR, the amount of target DNA doubles, allowing millions of copies to be produced from a single DNA molecule. PCR is a powerful tool used in various fields like forensics, medicine, and research.
The document summarizes the history and development of PCR technology. It describes key events like the concept of PCR being presented in 1984, the first PCR device called Mr. Cycle in 1985, the isolation of Taq polymerase in 1985 which enabled PCR without manual addition of reagents, and the licensing of PCR by Roche in 1991 for commercial use. It also provides details on the basic components and process of PCR.
Southern blotting and Northern blotting are techniques used to detect specific DNA and RNA sequences. Southern blotting involves transferring DNA fragments separated by gel electrophoresis to a membrane, then using a labeled probe to identify fragments by hybridization. Northern blotting follows similar steps but detects RNA, requiring formaldehyde treatment to denature RNA. Both techniques allow identification of specific molecules in complex mixtures.
Exploiting Artificial Intelligence for Empowering Researchers and Faculty, In...Dr. Vinod Kumar Kanvaria
Exploiting Artificial Intelligence for Empowering Researchers and Faculty,
International FDP on Fundamentals of Research in Social Sciences
at Integral University, Lucknow, 06.06.2024
By Dr. Vinod Kumar Kanvaria
Assessment and Planning in Educational technology.pptxKavitha Krishnan
In an education system, it is understood that assessment is only for the students, but on the other hand, the Assessment of teachers is also an important aspect of the education system that ensures teachers are providing high-quality instruction to students. The assessment process can be used to provide feedback and support for professional development, to inform decisions about teacher retention or promotion, or to evaluate teacher effectiveness for accountability purposes.
it describes the bony anatomy including the femoral head , acetabulum, labrum . also discusses the capsule , ligaments . muscle that act on the hip joint and the range of motion are outlined. factors affecting hip joint stability and weight transmission through the joint are summarized.
Macroeconomics- Movie Location
This will be used as part of your Personal Professional Portfolio once graded.
Objective:
Prepare a presentation or a paper using research, basic comparative analysis, data organization and application of economic information. You will make an informed assessment of an economic climate outside of the United States to accomplish an entertainment industry objective.
Thinking of getting a dog? Be aware that breeds like Pit Bulls, Rottweilers, and German Shepherds can be loyal and dangerous. Proper training and socialization are crucial to preventing aggressive behaviors. Ensure safety by understanding their needs and always supervising interactions. Stay safe, and enjoy your furry friends!
How to Build a Module in Odoo 17 Using the Scaffold MethodCeline George
Odoo provides an option for creating a module by using a single line command. By using this command the user can make a whole structure of a module. It is very easy for a beginner to make a module. There is no need to make each file manually. This slide will show how to create a module using the scaffold method.
বাংলাদেশের অর্থনৈতিক সমীক্ষা ২০২৪ [Bangladesh Economic Review 2024 Bangla.pdf] কম্পিউটার , ট্যাব ও স্মার্ট ফোন ভার্সন সহ সম্পূর্ণ বাংলা ই-বুক বা pdf বই " সুচিপত্র ...বুকমার্ক মেনু 🔖 ও হাইপার লিংক মেনু 📝👆 যুক্ত ..
আমাদের সবার জন্য খুব খুব গুরুত্বপূর্ণ একটি বই ..বিসিএস, ব্যাংক, ইউনিভার্সিটি ভর্তি ও যে কোন প্রতিযোগিতা মূলক পরীক্ষার জন্য এর খুব ইম্পরট্যান্ট একটি বিষয় ...তাছাড়া বাংলাদেশের সাম্প্রতিক যে কোন ডাটা বা তথ্য এই বইতে পাবেন ...
তাই একজন নাগরিক হিসাবে এই তথ্য গুলো আপনার জানা প্রয়োজন ...।
বিসিএস ও ব্যাংক এর লিখিত পরীক্ষা ...+এছাড়া মাধ্যমিক ও উচ্চমাধ্যমিকের স্টুডেন্টদের জন্য অনেক কাজে আসবে ...
Physiology and chemistry of skin and pigmentation, hairs, scalp, lips and nail, Cleansing cream, Lotions, Face powders, Face packs, Lipsticks, Bath products, soaps and baby product,
Preparation and standardization of the following : Tonic, Bleaches, Dentifrices and Mouth washes & Tooth Pastes, Cosmetics for Nails.
How to Add Chatter in the odoo 17 ERP ModuleCeline George
In Odoo, the chatter is like a chat tool that helps you work together on records. You can leave notes and track things, making it easier to talk with your team and partners. Inside chatter, all communication history, activity, and changes will be displayed.
2. Introduction to DNA
Fingerprinting and Forensics
Forensic science can be defined as the
intersection of law and science
First photography-then fingerprint- then, in
1985, DNA Fingerprinting
3. DNA Fingerprint
DNA fragments show unique patterns from one
person to the next.
Used in paternity disputes and as forensic
evidence.
4. Preparing a DNA Fingerprint
Specimen Collection- Could be a licked envelope, dirty
laundry, a cigarette butt, saliva
• Special precautions in handling specimens: gloves, disposable
instruments, avoid talking and sneezing, avoid touching sample
with your skin, air-dry the evidence before packaging so mold
does not grow
• Enemies of evidence: sunlight, high temperatures, bacteria,
moisture
• Ideal sample: 1 mL of fresh, whole blood (white blood cells)
treated with EDTA
5. RFLP
Restriction Fragment Length Polymorphism (RFLP)
• Nucleotide sequence variations in a region of DNA that
generates fragment length differences according to the
presence or absence of restriction enzyme recognition
sites.
9. Southern Blot
The first step in preparing a Southern Blot is to
cut genomic DNA and run on an agarose gel.
10. Southern Blot
The next step is to blot or transfer single stranded
DNA fragments on to a nylon membrane.
11. The next step is to hybridize a radioactively labeled DNA
probe to specific sequences on the membrane.
Southern Blot
12. Southern Blot
The last step is to expose the radioactively
labeled membrane to a large sheet of film.
You will only visualize bands where the probe
hybridized to the DNA.
17. PCR
Reaction requirements
• Template DNA – total genomic
DNA isolated from an organism
that contains a target region to be
amplified
• DNA primers - Short pieces of
single stranded DNA that flank the
target
• Taq DNA polymerase - Attaches
nucleotides on the growing strand
of DNA
• Nucleotides (GATC) – Polymerase
adds complementary nucleotides
to the template
18. PCR
Reactions are placed in a machine called a
thermal cycler. The machine cycles through
three temperatures.
19. PCR
1. Heat samples to 94°C for a minute or so to
denature the double stranded template DNA.
A little over a hundred years ago scientists discovered that patterns in the skin of the fingertips could be used to establish identity. In 1985 scientific researchers discovered a new kind of fingerprint. The unique signature in every person’s genetic makeup is called a DNA fingerprint. The uniform nature of DNA in a single individual and the genetic variability between individuals make DNA fingerprints useful to establish paternity and can also be used to implicate or exonerate a suspect in a crime.
Restriction Fragment Length Polymorphisms, or RFLPs for short, are generated due to mutations in recognition sites. Remember what a recognition site is? Correct! It’s the specific nucleotide sequence recognized by restriction enzymes. The restriction enzymes bind to the DNA and cut within the recognition site. If the nucleotide sequence has been mutated, then the restriction enzyme will not bind and therefore will not cut at that site. As this example shows, the change in the recognition site will produce different length fragments in a restriction digest. Individual one has two recognition sites for the EcoRI enzyme. That means EcoRI will cut this DNA fragment twice. What happens when you cut a piece of string twice? You get three pieces. The same is true when you cut linear DNA with molecular scissors like restriction enzymes. Two cuts generates three fragments labeled here as A, B, and C. In individual 2 the first recognition site is not present. Instead of GAATTC the sequence is now GAAATTC. EcoRI will not bind and cut at this location. That means in the same restriction digest, the restriction enzyme will only cut once and generate two fragments, D and C.
The fragments from the restriction digests can be separated by gel electrophoresis. Because the fragments are different lengths, they will run to different locations on a gel. This creates the unique RFLP pattern, or DNA fingerprint, for each individual.
Click on the RFLP animation to visualize the creation of a unique DNA fingerprint.
Gel electrophoresis is remarkably useful. One problem with gel electrophoresis is that it is not a permanent copy of the data. DNA continues to diffuse through the agarose matrix even after you stop the electrophoretic run. You can take a picture to keep a permanent record, but you cannot manipulate the DNA in any way. One way to create a permanent record of the restriction digest that can be tested and manipulated is to transfer and fix the DNA onto a nylon membrane. Edward Southern was the scientist who developed this technique, so it is now referred to as a “Southern Blot”. Human genomic DNA is 3 billion base pairs and contains thousands of recognition sites for restriction enzymes. When human genomic DNA is cut with restriction enzymes and run on an agarose gel you usually see a smear like the picture on the left. Transferring this DNA onto a nylon membrane allows researchers to hybridize specific probes to generate pictures like the one on the left. This picture is much clearer and more informative. Let’s learn how this southern blot hybridization is done.
As mentioned in the last slide, the first step in preparing a southern blot is to cut genomic DNA with restriction enzymes. The restriction enzymes will bind to and cut within it’s recognition site all along the 3 billion base pairs of genomic DNA. These restriction digests are then loaded onto an agarose gel and separated in an electrical field.
The DNA is then denatured in a basic solution. That means the hydrogen bonds holding the double stranded helix will be broken and the DNA will be single stranded. The single stranded DNA is then transferred onto a nylon membrane. The DNA is crosslinked onto the membrane with UV light or with heat. This creates a permanent copy of the DNA that was separated on the agarose gel.
You may have wondered why the DNA had to be made single stranded before it was transferred to the membrane. Well, that was so you could hybridize a probe to the transferred DNA. A DNA probe is a short, single stranded molecule that is labeled (either radioactively or fluorescently). Because both the probe and the transferred DNA are single stranded, when incubated together they complementarily base pair. When the probe binds to it’s complement on the DNA it is called hybridizing the probe.
Because the probe is radioactively labeled, it can be exposed onto a large sheet of x-ray film. After exposure to the film you will not see the long smears of genomic DNA, you will only be able to visualize the bands where the probe hybridized to the DNA on the membrane.
Click on the Southern Blot Animation to visualize this process.
Another reason why every individual has a unique DNA fingerprint is because of regions called variable number tandem repeats, or VNTR’s. A VNTR is a sequence that is repeated multiple times. The number of repeats varies from person to person. This example shows the sequence GATC repeated 5 times in one individual and only 2 times in a second individual.
These VNTR regions usually occur in introns. Do you remember what those are? Yes, the are noncoding regions of DNA that are cut out before the mature mRNA leaves the nucleus to make protein. That means the number of repeats in the VNTR region will have no effect on protein expression and therefore no effect on phenotypic traits. Because of this, VNTR loci are not under the same evolutionary selective pressure and are therefore highly variable.
You can visualize the VNTR regions by amplifying them with a technique called polymerase chain reaction, or PCR. The PCR reactions can be run on a gel such as the one pictured here. This gel shows the amplification of a common VNTR locus called the DS180 repeat found in chromosome 16. The DS180 repeat can be used to trace the ancestry of an individual within a species. More on genealogy later, but now let’s learn how to do a PCR reaction.
PCR was developed in the mid 1980’s by Kerry Mullis and it is really just DNA replication in a tube. So the requirements for a PCR reaction are the same as the requirements for DNA replication. You must add the template DNA, which can often be total genomic DNA. The target is the region on the template that you wish to amplify. You need to add primers, which are short single stranded molecules of DNA that flank the target region. Just like in replication, the primers allow DNA polymerase to attach and begin replication. A special DNA polymerase is added, called Taq DNA polymerase. This polymerase acts just like any other DNA polymerase in that it attaches nucleotides to the newly growing strand of DNA by matching the complementary nucleotide on the template strand. Taq DNA polymerase is special because it was discovered in a bacteria that grows in a hot spring. That means that it can withstand extremely high temperatures. We’ll explain why that is important in just a minute. If Taq DNA polymerase is going to extend the growing strand of DNA, there must be deoxynucleotides in the reaction mix. These are the G’s, A’s, T’s and C’s that polymerase incorporates into the new strand.
Once all of the components have been added to the reaction tube, the reactions are placed in a machine called a thermal cycler. The thermal cycler is an instrument that is capable of rapidly changing temperature over very short time intervals. The machine repeatedly cycles through three different temperatures during a PCR reaction. Let’s learn about these temperature cycles.
The first temperature is 94 degrees celsius. This is called the denaturation step because the hydrogen bonds between the double stranded DNA are broken, or denatured, making all of the DNA single stranded.
The temperature is then dropped to around 55 degrees celsius. This is called the annealing step because it allows the single stranded primers to find and attach to their complement on the template DNA. Remember that the primers were created to specifically flank the target region you desired to amplify.
The third step is to raise the temperature to 72 degrees celsius. This step is called the elongation step because at this temperature the Taq DNA polymerase is extending from the primer by adding nucleotides that are complementary to the template to the newly growing strand of DNA. At the end of this third temperature step, you have completely replicated the target region of DNA and that means you have doubled the number of copies of the target.
The thermal cycler literally cycles through these three temperature steps for the number of times programmed into the machine, usually around 30 times.
Every time the machine cycles through the temperatures, the number of copies of the target is doubled. That means after one cycle you will have two copies, then four copies, then 8, 16, 32….and so on. Because of this logarithmic amplification, PCR is an easy way to generate lots of copies of a desired region of DNA and has numerous applications in biotechnology.
Click on the PCR animation to visualize this process.
Let’s talk about some applications of RFLP and VNTR analysis. One application for the medical field is diagnosing disease. Sickle cell anemia is a disease that is characterized by red blood cells that assume an abnormal, rigid, sickle shape. These abnormally shaped red blood cells cannot properly bind and transport oxygen in the blood. As you may recall from module one, this disease is the result of a single point mutation that causes the amino acid valine to replace glutamic acid in the amino acid chain. This mutation also creates an additional recognition site for the restriction enzyme DdeI. RFLP analysis will reflect this difference, as you can see in the picture of the gel, and can be used to diagnose sickle cell patients or alert potential parents that they are carriers of the disease.
In addition to diagnosing a number of different diseases, RFLP analysis can be used for disputed paternity suits. DNA from the mother, child, and potential fathers are received in the form of blood samples or cheek swabs. Once the DNA has been isolated, RFLP analysis can be done to determine the paternity of the child. This is possible because children receive their DNA only from their mother or father, that means that any bands a child has must also be represented by one parent or the other. In this example, the highlighted bands that did not come from the mother must have come from the father. You can see by examining the data that father 2 matches every band with the child that did not come from the mother. Therefore father 1 can be excluded and father 2 can be identified as this child’s biological father. Every year roughly 250,000 paternity suits are filed in the United States. Using RFLP analysis, verifying a child’s parentage and resolving child support or custody disputes is relatively easy.
And of course RFLP and VNTR analysis can be used as forensic evidence. A crime scene is full of sources of DNA evidence including dirty laundry, a licked envelope, or a cigarette butt. Tiny blood stains, a smear of dried semen, or a trace of saliva is often all it takes to crack a case. In this example, a woman was found murdered in her apartment. The crime scene investigator collected samples of the woman’s blood, samples from the knife found under the body, and samples of blood from underneath the woman’s fingernails. The police suspected her ex-boyfriend was the perpetrator of the crime. A search of his apartment turned up a bloody shirt that he claimed was the result of falling into a rose bush while gardening. The evidence was very incriminating, so the investigator took him in for further questioning took a blood sample from him. This gel is a copy of the data obtained by PCR of VNTR loci in the labeled samples. You can see by matching the bands that the blood on the knife belonged to the woman and the blood under the woman’s fingernails belonged to her and another suspect that does not match the ex-boyfriend. Furthermore, the analysis of the blood stains on the ex-boyfriend’s shirt seem to confirm his alibi because his is the only blood on the shirt. Without DNA analysis, this man could have been convicted for this crime. Instead, he was exonerated and the real perpetrator was brought to justice. CODIS is a DNA database funded by the FBI that stores DNA profiles of convicted felons created by federal, state, and local crime labs. The standard in DNA profiles created for forensic evidence are to use 13 markers plus one to determine sex. If any two samples have matching genotypes at all 13 CODIS loci, it is a virtual certainty that the two DNA samples came from the same individual (or an identical twin). The applications for RFLP and VNTR loci extend beyond the scope of this module, but as you can see from these few examples this technology is powerful and necessary.
Another application of analysis of RFLP and VNTR loci in establishing familial relationships is in the area of molecular genealogy. Traditional genealogy involves extensive search of historical records. But our DNA also contains a record of our familial relationships. DNA analysis can provide clues about what part of the world our ancestors came from. We learned in module one about different types of mutations. As DNA is copied and passed down through generations, it gradually accumulates more mutations. These mutations account for human genetic variation. RFLP and VNTR analysis of these mutations show that people who are more closely related will share more similarities in their DNA. These individuals are said to have the same haplotype. The number of differences between your haplotype and the haplotype of another person can tell you the number of generations you have to go back to find a common ancestor. Click on the genealogy animation to learn more about molecular genealogy.