Southern blotting
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Southern blotting is a technique used to detect specific DNA sequences in a sample. It involves extracting DNA from a source, cutting it into fragments using restriction enzymes, separating the fragments by size through gel electrophoresis, and then transferring the fragments to a membrane and using a radioactive probe to detect sequences that hybridize. Southern blotting is used for gene discovery, mapping, and confirming that a known sequence has been incorporated into the genome of a genetically modified organism.
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Southern blot
Southern Blot is a technique invented by Edwin Southern in the 1970s to locate specific DNA sequences within a larger sample of DNA. It works by cutting DNA with restriction enzymes, separating the fragments via gel electrophoresis, transferring them to a membrane, and using a radioactive or enzyme-labeled probe to bind to complementary DNA sequences, revealing their locations. Southern Blot can be used to map genes, for forensic analysis, and to discover new genes or detect diseases.
Southern blotting
Southern blotting is a technique used to detect specific DNA sequences. It involves separating DNA fragments by size via gel electrophoresis, transferring them to a membrane, and using a labeled probe to hybridize to the target sequence. The probe binds only to complementary DNA fragments, allowing their detection and location on the membrane via autoradiography. Key steps include restriction enzyme digestion of DNA, gel electrophoresis, membrane transfer, probe hybridization, and x-ray film development to visualize hybridized fragments. Southern blotting is used for applications like mutation detection, gene rearrangement studies, and forensic analysis.
Southern blot power point
Southern blotting is a DNA analysis technique developed by Edwin Southern in 1975 that involves separating DNA fragments by size using gel electrophoresis, transferring them to a membrane, and using a radioactive probe to detect complementary DNA sequences by their position on the membrane after developing an X-ray film. Southern blots are commonly used to verify that a desired DNA sequence has been successfully inserted into the genome of a genetically modified organism.
DNA Fingerprinting
DNA fingerprinting is a technique used to identify individuals based on their unique DNA patterns. It involves isolating DNA from a sample, cutting the DNA into fragments of varying lengths, sorting the fragments by size, and then probing the fragments to produce a pattern - the DNA fingerprint - that can be used to identify an individual. DNA fingerprinting has various uses, including diagnosing inherited disorders, linking suspects to biological evidence in criminal cases, and personal identification such as for military personnel or paternity tests.
Dna fingerprinting- Logan Briggs
DNA fingerprinting is a process used to identify individuals by their unique DNA patterns. It involves cutting DNA into fragments using restriction enzymes, separating the fragments via gel electrophoresis, transferring them to a membrane, and detecting specific DNA sequences using radioactive probes. When exposed to X-ray film, the probe-hybridized fragments produce a pattern of bands that serves as an individual's unique DNA fingerprint. DNA fingerprinting has applications in criminal investigations, genetic trait and disease analysis, animal breeding, and disease tracking and origin identification.
Southern blot kanchana
Southern blotting is a technique used to detect specific DNA sequences. It involves purifying DNA from cells, cutting the DNA into fragments using restriction enzymes, separating the fragments via gel electrophoresis, transferring the fragments to a membrane, and using a labeled probe to identify fragments that are complementary to the probe. The final step is to visualize the bound probes on the membrane through autoradiography to determine which DNA fragments are present in the sample. Southern blotting can be used for applications like DNA fingerprinting, paternity testing, and identifying genetic mutations or infectious agents.
Blotting techniques1
This document discusses different blotting techniques used to detect DNA, RNA, and proteins. Southern blotting is used to detect DNA and was developed by Edwin Southern in 1975. It involves separating DNA fragments by size, transferring them to a membrane, and using a probe to detect specific sequences. Northern blotting detects RNA and was developed in 1979. It similarly separates RNA and uses a probe to detect sequences of interest. Western blotting detects proteins and was developed in 1981. It separates proteins by SDS-PAGE gel electrophoresis, transfers them to a membrane, and uses an antibody probe to detect the target protein. These techniques are widely used in research and diagnostics.
NEXT GENERATION SEQUENCING
This presentation is explains about the genome sequencing, its traditional method and modern method. This basically focus on Next Generation Sequencing and its types.
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Southern blot
Southern Blot is a technique invented by Edwin Southern in the 1970s to locate specific DNA sequences within a larger sample of DNA. It works by cutting DNA with restriction enzymes, separating the fragments via gel electrophoresis, transferring them to a membrane, and using a radioactive or enzyme-labeled probe to bind to complementary DNA sequences, revealing their locations. Southern Blot can be used to map genes, for forensic analysis, and to discover new genes or detect diseases.
Southern blotting
Southern blotting is a technique used to detect specific DNA sequences. It involves separating DNA fragments by size via gel electrophoresis, transferring them to a membrane, and using a labeled probe to hybridize to the target sequence. The probe binds only to complementary DNA fragments, allowing their detection and location on the membrane via autoradiography. Key steps include restriction enzyme digestion of DNA, gel electrophoresis, membrane transfer, probe hybridization, and x-ray film development to visualize hybridized fragments. Southern blotting is used for applications like mutation detection, gene rearrangement studies, and forensic analysis.
Southern blot power point
Southern blotting is a DNA analysis technique developed by Edwin Southern in 1975 that involves separating DNA fragments by size using gel electrophoresis, transferring them to a membrane, and using a radioactive probe to detect complementary DNA sequences by their position on the membrane after developing an X-ray film. Southern blots are commonly used to verify that a desired DNA sequence has been successfully inserted into the genome of a genetically modified organism.
DNA Fingerprinting
DNA fingerprinting is a technique used to identify individuals based on their unique DNA patterns. It involves isolating DNA from a sample, cutting the DNA into fragments of varying lengths, sorting the fragments by size, and then probing the fragments to produce a pattern - the DNA fingerprint - that can be used to identify an individual. DNA fingerprinting has various uses, including diagnosing inherited disorders, linking suspects to biological evidence in criminal cases, and personal identification such as for military personnel or paternity tests.
Dna fingerprinting- Logan Briggs
DNA fingerprinting is a process used to identify individuals by their unique DNA patterns. It involves cutting DNA into fragments using restriction enzymes, separating the fragments via gel electrophoresis, transferring them to a membrane, and detecting specific DNA sequences using radioactive probes. When exposed to X-ray film, the probe-hybridized fragments produce a pattern of bands that serves as an individual's unique DNA fingerprint. DNA fingerprinting has applications in criminal investigations, genetic trait and disease analysis, animal breeding, and disease tracking and origin identification.
Southern blot kanchana
Southern blotting is a technique used to detect specific DNA sequences. It involves purifying DNA from cells, cutting the DNA into fragments using restriction enzymes, separating the fragments via gel electrophoresis, transferring the fragments to a membrane, and using a labeled probe to identify fragments that are complementary to the probe. The final step is to visualize the bound probes on the membrane through autoradiography to determine which DNA fragments are present in the sample. Southern blotting can be used for applications like DNA fingerprinting, paternity testing, and identifying genetic mutations or infectious agents.
Blotting techniques1
This document discusses different blotting techniques used to detect DNA, RNA, and proteins. Southern blotting is used to detect DNA and was developed by Edwin Southern in 1975. It involves separating DNA fragments by size, transferring them to a membrane, and using a probe to detect specific sequences. Northern blotting detects RNA and was developed in 1979. It similarly separates RNA and uses a probe to detect sequences of interest. Western blotting detects proteins and was developed in 1981. It separates proteins by SDS-PAGE gel electrophoresis, transfers them to a membrane, and uses an antibody probe to detect the target protein. These techniques are widely used in research and diagnostics.
NEXT GENERATION SEQUENCING
This presentation is explains about the genome sequencing, its traditional method and modern method. This basically focus on Next Generation Sequencing and its types.
Gibson Assembly: a seamless molecular cloning technique
This is an Open Educational Resource (OER) coursework for my Digital Education module. This OER compares and contrasts conventional restriction-based cloning with modern Gibson Assembly. The graphics in the Gibson Assembly sections can be easily reused and customized with freeware such as Inkscape to construct high-resolution PowerPoint animation in Molecular Cloning lecture.
Blotting techniques
This document discusses various blotting techniques used to detect DNA, RNA, and proteins, including Southern blotting, Northern blotting, and Western blotting. It provides detailed descriptions of the procedures for each technique, including separating biomolecules by electrophoresis, transferring them to a membrane, hybridizing probes, and detecting bound probes. The techniques allow visualization and analysis of specific biomolecules within complex samples.
Blotting techniques ppt
The document discusses various blotting techniques used to detect DNA, RNA, and proteins. It describes Southern blotting for detecting DNA, Northern blotting for detecting RNA, and Western blotting for detecting proteins. It provides details on the steps involved in each technique, including separating biomolecules by electrophoresis, transferring them to a membrane, and using probes for detection through hybridization or antibody binding.
Nucleic acid hybridization
Nucleic acid hybridization is a technique used to identify specific DNA sequences. It involves denaturing DNA or RNA samples and probes, followed by annealing of the probes to complementary sequences. There are two main types: Southern blotting separates DNA fragments by gel electrophoresis before hybridization with probes, while Northern blotting separates RNA this way. Both techniques allow detection of specific sequences through the use of labeled probes.
DNA Sequencing
The document discusses DNA sequencing, including its history, common methods like Sanger sequencing, the steps involved, challenges and limitations. DNA sequencing determines the order of nucleotides in DNA and has various applications such as identifying genetic diseases, forensics, and studying genome evolution. Sanger sequencing, developed in 1977, works by using chain-terminating dideoxynucleotides during DNA replication to determine the sequence.
Blotting techniques (manish)
The document discusses various blotting techniques used to transfer and detect DNA, RNA, and proteins, including Southern blotting for DNA, Northern blotting for RNA, and Western blotting for proteins. Southern blotting involves separating DNA fragments via gel electrophoresis, transferring them to a membrane, and using a probe to detect specific sequences. It allows researchers to identify genes, map genomes, and study evolution and disease.
Gibson Assembly in Cloning
Gibson assembly is a DNA assembly method that allows joining of multiple DNA fragments in a single isothermal reaction to produce a single double stranded molecule. It involves three enzymes - T5 exonuclease, DNA phusion polymerase, and Taq DNA ligase. The enzymes are mixed with DNA fragments that have 20-40 bp overlapping regions. T5 exonuclease exposes complementary sequences, polymerase fills gaps, and ligase seals nicks to produce the full assembly. Gibson assembly has advantages like being carried out in one tube in one step with no restriction enzymes, but requires long overlapping oligonucleotides and errors can occur at junctions.
DNA Fingerprinting or DNA Profiling
DNA Fingerprinting involves different stages -
1. First is isolation of DNA from target cell.
2. Then treatment of restriction digestion of DNA to generate DNA fragments.
3. Then separation of DNA fragments using Agarose Gel Electrophoresis .
4. Transfer of Agarose Gel DNA bands to nitrocellulose membrane filter. This filter paper after the DNA transfer is known as Southern Blot .
5. Then use of complementary DNA Probe to bind with target DNA .
6. X- Ray film exposure which represent Dark bands of DNA called DNA Fingerprints .
Southern blotting
these slides can easily give better understanding to the reader about the Steps involved in Southern Blotting.
Dna fingerprinting
DNA fingerprinting is a technique used to identify individuals based on their unique DNA patterns. It involves isolating DNA from a sample, cutting the DNA into fragments of varying lengths using restriction enzymes, sorting the fragments by size through gel electrophoresis, transferring the fragments to a nylon membrane, and probing the membrane to produce a pattern - the DNA fingerprint - that can be used to identify an individual. DNA fingerprinting has uses in personal identification such as identifying casualties or missing military personnel.
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Strawberry dn aextraction
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DND sequencing
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DNA FINGERPRINTING
DNA fingerprinting is a method used to identify individuals based on variations in their DNA sequences. It involves using restriction enzymes to cut DNA into fragments at specific sequences, which are then separated by size using gel electrophoresis. The pattern of fragment sizes is unique to each individual and can be used to identify a person, determine paternity, solve criminal cases, diagnose genetic disorders, and develop cures. The document provides details on the different types of tandem repeats like satellites, minisatellites, and microsatellites that contribute to variations between individuals, as well as the basic process and applications of DNA fingerprinting.
Southern Blotting (SB) 4 jan 2015 final
The power Point presentation contains 38 slides explaining about different steps involved in Southern Blotting such as DNA Isolation, Restriction digestion, Separation of DNA fragments by gel electrophoresis, denaturation of Double stranded DNA , transfer of fragments from gel to membrane ( blotting) , hybridization and detection by autoradiography. Applications of Southern blotting have also been discussed
Northern blotting ppt
Northern blotting is a technique used to detect specific mRNAs in a sample. It involves separating RNA fragments by electrophoresis, transferring them to a membrane, then using a probe with a complementary nucleotide sequence to identify particular mRNA transcripts through hybridization. This allows researchers to determine which genes are actively being expressed and study gene expression under different conditions. The key steps are isolating RNA, separating fragments by size in a gel, immobilizing them on a membrane, then incubating with a labeled probe and detecting hybridized bands through autoradiography or other methods. Northern blotting provides a simple way to analyze mRNA levels and splicing variations.
What is blotting
Blotting techniques such as Southern blotting, Northern blotting, and Western blotting allow researchers to detect specific DNA, RNA, and protein sequences by transferring them from a gel to a membrane and using probes to identify the targets. Southern blotting is used to detect DNA, Northern blotting detects RNA, and Western blotting identifies proteins. These techniques play important roles in research areas like gene expression analysis, disease diagnosis, forensics, and more by providing sensitive and specific detection of biomolecules.
Southern Blotting Technique
This document describes the process of Southern blotting, which is used to detect specific DNA sequences in a sample. It involves purifying DNA from cells, cutting the DNA into fragments using restriction enzymes, separating the fragments by size via gel electrophoresis, transferring them to a membrane, then using a radioactive probe to detect fragments that hybridize based on complementary base pairing. The technique allows identification of mutations, deletions and other genetic variations, and has applications in medical diagnosis, forensics and other areas requiring DNA analysis.
DNA Fingerprinting
DNA Fingerprinting Explained, Techniques Used, Usage, Limitations and Contradictions.
*I won an Award for the Best Power Point Project Presentation in class 12th for this project. :D
southern blotting (used to detect the presence of a particular piece of DNA i...
The technique was developed by E.M. Southern in 1975.
The Southern blot is used to detect the presence of a particular piece of DNA in a sample
mujahid hussain, Department of Botany, University of Sargodha, Sargodha, Punjab, Pakistan
Southern blotting
Southern blotting is a technique used to detect specific DNA sequences. It involves extracting DNA from cells, cutting the DNA with restriction enzymes, separating fragments via electrophoresis, transferring DNA to a membrane, then using a labeled probe to hybridize and identify the target sequence via autoradiography. Southern blotting allows identification of mutations, deletions and rearrangements, and is used for cancer prognosis, genetic disease diagnosis, DNA fingerprinting and more. It is an effective detection method but is also complex, labor-intensive, and time-consuming.
Southern blotting
Southern blotting is a technique used to detect specific DNA sequences within DNA samples. It involves digesting DNA samples with restriction enzymes, separating the fragments via gel electrophoresis, transferring the fragments to a membrane, and using radioactive probes to hybridize to complementary DNA sequences, which are then visualized. The key steps are digesting DNA, separating fragments by size via gel electrophoresis, transferring fragments to a membrane, hybridizing with radioactive probes, and detecting hybridized probes.
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This is an Open Educational Resource (OER) coursework for my Digital Education module. This OER compares and contrasts conventional restriction-based cloning with modern Gibson Assembly. The graphics in the Gibson Assembly sections can be easily reused and customized with freeware such as Inkscape to construct high-resolution PowerPoint animation in Molecular Cloning lecture.
Blotting techniques
This document discusses various blotting techniques used to detect DNA, RNA, and proteins, including Southern blotting, Northern blotting, and Western blotting. It provides detailed descriptions of the procedures for each technique, including separating biomolecules by electrophoresis, transferring them to a membrane, hybridizing probes, and detecting bound probes. The techniques allow visualization and analysis of specific biomolecules within complex samples.
Blotting techniques ppt
The document discusses various blotting techniques used to detect DNA, RNA, and proteins. It describes Southern blotting for detecting DNA, Northern blotting for detecting RNA, and Western blotting for detecting proteins. It provides details on the steps involved in each technique, including separating biomolecules by electrophoresis, transferring them to a membrane, and using probes for detection through hybridization or antibody binding.
Nucleic acid hybridization
Nucleic acid hybridization is a technique used to identify specific DNA sequences. It involves denaturing DNA or RNA samples and probes, followed by annealing of the probes to complementary sequences. There are two main types: Southern blotting separates DNA fragments by gel electrophoresis before hybridization with probes, while Northern blotting separates RNA this way. Both techniques allow detection of specific sequences through the use of labeled probes.
DNA Sequencing
The document discusses DNA sequencing, including its history, common methods like Sanger sequencing, the steps involved, challenges and limitations. DNA sequencing determines the order of nucleotides in DNA and has various applications such as identifying genetic diseases, forensics, and studying genome evolution. Sanger sequencing, developed in 1977, works by using chain-terminating dideoxynucleotides during DNA replication to determine the sequence.
Blotting techniques (manish)
The document discusses various blotting techniques used to transfer and detect DNA, RNA, and proteins, including Southern blotting for DNA, Northern blotting for RNA, and Western blotting for proteins. Southern blotting involves separating DNA fragments via gel electrophoresis, transferring them to a membrane, and using a probe to detect specific sequences. It allows researchers to identify genes, map genomes, and study evolution and disease.
Gibson Assembly in Cloning
Gibson assembly is a DNA assembly method that allows joining of multiple DNA fragments in a single isothermal reaction to produce a single double stranded molecule. It involves three enzymes - T5 exonuclease, DNA phusion polymerase, and Taq DNA ligase. The enzymes are mixed with DNA fragments that have 20-40 bp overlapping regions. T5 exonuclease exposes complementary sequences, polymerase fills gaps, and ligase seals nicks to produce the full assembly. Gibson assembly has advantages like being carried out in one tube in one step with no restriction enzymes, but requires long overlapping oligonucleotides and errors can occur at junctions.
DNA Fingerprinting or DNA Profiling
DNA Fingerprinting involves different stages -
1. First is isolation of DNA from target cell.
2. Then treatment of restriction digestion of DNA to generate DNA fragments.
3. Then separation of DNA fragments using Agarose Gel Electrophoresis .
4. Transfer of Agarose Gel DNA bands to nitrocellulose membrane filter. This filter paper after the DNA transfer is known as Southern Blot .
5. Then use of complementary DNA Probe to bind with target DNA .
6. X- Ray film exposure which represent Dark bands of DNA called DNA Fingerprints .
Southern blotting
these slides can easily give better understanding to the reader about the Steps involved in Southern Blotting.
Dna fingerprinting
DNA fingerprinting is a technique used to identify individuals based on their unique DNA patterns. It involves isolating DNA from a sample, cutting the DNA into fragments of varying lengths using restriction enzymes, sorting the fragments by size through gel electrophoresis, transferring the fragments to a nylon membrane, and probing the membrane to produce a pattern - the DNA fingerprint - that can be used to identify an individual. DNA fingerprinting has uses in personal identification such as identifying casualties or missing military personnel.
Graph Algorithms: Breadth-First Search (BFS)
This document describes breadth-first search (BFS) graph algorithms. It begins by defining graphs and different graph representations. It then explains BFS, which explores a graph by exploring all of the neighbor nodes of the starting node, then all of the neighbors of the neighbors, and so on outwards in "waves" or breadth-first manner. The document provides pseudocode for BFS and walks through an example run on a sample graph to demonstrate how BFS works. It concludes by discussing the running time of BFS and providing an example programming assignment for implementing BFS.
Strawberry dn aextraction
This document provides instructions for extracting and viewing DNA from a strawberry. It involves mashing a strawberry in a bag with a DNA extracting solution, then pouring the mixture through a filter and into a test tube. A few drops of alcohol are added, which causes the DNA to become visible as a white cloud forming at the interface between the two liquids. The document also provides background information on what DNA is, its structure, and how much DNA is shared between humans and other organisms.
DND sequencing
This document defines DNA sequencing and describes some common DNA sequencing methods. It explains that DNA sequencing determines the order of the four nucleotide bases that make up DNA. It then describes two basic DNA sequencing methods - Maxam-Gilbert chemical sequencing and Sanger chain termination sequencing. For Sanger sequencing, it provides details on how fluorescent dideoxynucleotides are used to randomly terminate DNA strands during replication, allowing the sequence to be read from the resulting fragments.
DNA FINGERPRINTING
DNA fingerprinting is a method used to identify individuals based on variations in their DNA sequences. It involves using restriction enzymes to cut DNA into fragments at specific sequences, which are then separated by size using gel electrophoresis. The pattern of fragment sizes is unique to each individual and can be used to identify a person, determine paternity, solve criminal cases, diagnose genetic disorders, and develop cures. The document provides details on the different types of tandem repeats like satellites, minisatellites, and microsatellites that contribute to variations between individuals, as well as the basic process and applications of DNA fingerprinting.
Southern Blotting (SB) 4 jan 2015 final
The power Point presentation contains 38 slides explaining about different steps involved in Southern Blotting such as DNA Isolation, Restriction digestion, Separation of DNA fragments by gel electrophoresis, denaturation of Double stranded DNA , transfer of fragments from gel to membrane ( blotting) , hybridization and detection by autoradiography. Applications of Southern blotting have also been discussed
Northern blotting ppt
Northern blotting is a technique used to detect specific mRNAs in a sample. It involves separating RNA fragments by electrophoresis, transferring them to a membrane, then using a probe with a complementary nucleotide sequence to identify particular mRNA transcripts through hybridization. This allows researchers to determine which genes are actively being expressed and study gene expression under different conditions. The key steps are isolating RNA, separating fragments by size in a gel, immobilizing them on a membrane, then incubating with a labeled probe and detecting hybridized bands through autoradiography or other methods. Northern blotting provides a simple way to analyze mRNA levels and splicing variations.
What is blotting
Blotting techniques such as Southern blotting, Northern blotting, and Western blotting allow researchers to detect specific DNA, RNA, and protein sequences by transferring them from a gel to a membrane and using probes to identify the targets. Southern blotting is used to detect DNA, Northern blotting detects RNA, and Western blotting identifies proteins. These techniques play important roles in research areas like gene expression analysis, disease diagnosis, forensics, and more by providing sensitive and specific detection of biomolecules.
Southern Blotting Technique
This document describes the process of Southern blotting, which is used to detect specific DNA sequences in a sample. It involves purifying DNA from cells, cutting the DNA into fragments using restriction enzymes, separating the fragments by size via gel electrophoresis, transferring them to a membrane, then using a radioactive probe to detect fragments that hybridize based on complementary base pairing. The technique allows identification of mutations, deletions and other genetic variations, and has applications in medical diagnosis, forensics and other areas requiring DNA analysis.
DNA Fingerprinting
DNA Fingerprinting Explained, Techniques Used, Usage, Limitations and Contradictions.
*I won an Award for the Best Power Point Project Presentation in class 12th for this project. :D
southern blotting (used to detect the presence of a particular piece of DNA i...
The technique was developed by E.M. Southern in 1975.
The Southern blot is used to detect the presence of a particular piece of DNA in a sample
mujahid hussain, Department of Botany, University of Sargodha, Sargodha, Punjab, Pakistan
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Gibson Assembly: a seamless molecular cloning technique
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Southern blotting
Southern blotting is a technique used to detect specific DNA sequences. It involves extracting DNA from cells, cutting the DNA with restriction enzymes, separating fragments via electrophoresis, transferring DNA to a membrane, then using a labeled probe to hybridize and identify the target sequence via autoradiography. Southern blotting allows identification of mutations, deletions and rearrangements, and is used for cancer prognosis, genetic disease diagnosis, DNA fingerprinting and more. It is an effective detection method but is also complex, labor-intensive, and time-consuming.
Southern blotting
Southern blotting is a technique used to detect specific DNA sequences within DNA samples. It involves digesting DNA samples with restriction enzymes, separating the fragments via gel electrophoresis, transferring the fragments to a membrane, and using radioactive probes to hybridize to complementary DNA sequences, which are then visualized. The key steps are digesting DNA, separating fragments by size via gel electrophoresis, transferring fragments to a membrane, hybridizing with radioactive probes, and detecting hybridized probes.
Southern blot method
The document describes the Southern blot technique, which was discovered in 1973 by Edwin Southern at Edinburgh University. The Southern blot allows for the detection of specific DNA sequences and involves separating DNA fragments using gel electrophoresis, transferring them to a membrane, incubating the membrane with a probe that binds to complementary DNA sequences, and revealing the location of the probe using a colorless substrate and X-ray film.
Blotting techniques
This document describes three types of blotting techniques - Southern blotting, Northern blotting, and Western blotting. Southern blotting is used to detect DNA fragments separated by agarose gel electrophoresis. Northern blotting detects specific RNA sequences separated by gel electrophoresis. Western blotting identifies proteins separated by SDS-PAGE gel using an antibody probe. The document provides detailed procedures and applications for each type of blotting.
Southern hybridization
Southern blotting is a technique used to detect specific DNA sequences in a DNA sample. It involves extracting DNA from cells, cutting the DNA into fragments using restriction enzymes, separating the fragments via gel electrophoresis, transferring the DNA fragments to a membrane, and using a labeled probe to detect fragments that are complementary to the probe through hybridization. Southern blotting is useful for identifying mutations, DNA fingerprinting, and detecting DNA in applications like prenatal screening and forensics. While effective for detecting specific DNA sequences, it is a complex, time-consuming, and labor-intensive technique.
Heterochromatin and euchromatin mains
This document discusses the structures and functions of heterochromatin and euchromatin. Heterochromatin is tightly packed and transcriptionally inactive, found near centromeres and telomeres. Euchromatin is loosely packed and contains most actively transcribed genes. The basic unit of DNA packing is the nucleosome, which involves DNA wound around histone proteins. Heterochromatin and euchromatin differ in their genetic activity, location within chromosomes, and condensation levels during interphase.
Chromatin and chromosomes
Chromatin is the complex of DNA and proteins found in the eukaryotic nucleus. Chromatin is packaged and organized into chromosomes. DNA is wrapped around histone proteins forming nucleosomes to efficiently package the DNA into chromosomes, which make up the full genome of an organism.
Northern blotting
Northern blotting is a technique used to detect specific RNA sequences in a sample. It involves separating RNA samples by size through gel electrophoresis, transferring them to a nylon membrane, and using a labeled probe to detect the target RNA sequence through hybridization. The target RNA will be visible as a band on the membrane where the probe has bound. This allows researchers to observe gene expression levels under different conditions like development or disease.
Rna splicing
RNA splicing is the process by which introns, or non-coding sequences, are removed from pre-messenger RNA (pre-mRNA) to produce mature mRNA that can be translated into protein. Most genes contain introns that are removed by a spliceosome, a complex of RNA and proteins, leaving just the coding exons to form mRNA. Alternative splicing allows one gene to encode multiple proteins by selecting different combinations of exons. Errors in splicing can cause diseases if they result in truncated or abnormal proteins.
Chromatin structure "DNA+CHROMOSOME"
Chromatin structure "dna with histones "
and genes +regulation of transcription by chromatin structure
chromosome and cell cycle
STRUCTURE AND ORGANIZATION OF CHROMATIN
This document provides information on the structure and organization of chromatin. It discusses three main levels of chromatin organization:
1) Nucleosomes, which involve DNA wrapping around histone proteins in "beads on a string" structures.
2) The 30nm chromatin fiber, where nucleosomes are further compacted into a solenoid structure through histone H1 binding.
3) Higher-order compaction into metaphase chromosomes, where DNA is organized into loops anchored to a protein scaffold.
The document also describes different chromatin types (euchromatin and heterochromatin), chromatin composition, and models of chromatin structure including the nucleosome model.
Chromatin
1st ChIP-SAGE uses ChIP to purify chromatin, then crosslinks are reversed and linkers are ligated to DNA ends before digestion with NlaIII and MmeI to produce 21-22 bp sequence tags for cloning and sequencing.
2nd Deposition of histone variants like H2A.Z and H3.3 mark boundaries of regulatory regions in genomes and create hierarchical nucleosome stability.
3rd Interpreting ChIP experiments requires considering antibody specificity, chromatin preparation methods, and that results reflect average modification states across cells.
SPLICING
The document discusses RNA splicing and its importance. It covers how splicing removes introns and joins exons to create mature RNA for protein production. Alternative splicing can generate multiple protein isoforms from a single gene with different functions. Myotonic dystrophy type 1 is discussed as an example of a disorder caused by aberrant splicing, changing the form of the PKM2 enzyme critical for muscle cell metabolism. A new complex called the Little Elongation Complex is revealed to play a key role in transcription of small nuclear RNAs required for splicing. Understanding splicing machinery could open doors to novel disease treatment approaches.
Southern blot
The document summarizes the Southern blot technique. It involves digesting DNA with restriction enzymes, separating fragments by size via gel electrophoresis, transferring fragments to a membrane, and detecting a specific DNA sequence using a radiolabeled probe. The Southern blot allows determining the size of a restriction fragment, measuring amounts between samples, and locating a specific sequence within a complex DNA mixture. It has applications in gene discovery, mapping, identification of transgenic genes, and DNA fingerprinting.
Northern blotting
Northern blotting (RNA Dectection by botting process)
Southern and northern blotting comparison (similarities and differences)
DNA sequencing by OLAGBAYE B.ABEL
DNA sequencing methods determine the order of nucleotide bases in a DNA sample. The first methods were developed in the 1970s by Maxam and Gilbert (chemical method) and Sanger (chain termination method). The Sanger method is now most commonly used. It involves DNA synthesis with chain terminating nucleotides to generate fragments of different lengths that can be separated and read to determine the DNA sequence. DNA sequencing has revolutionized biological sciences by allowing for gene mapping and identification of genetic variations linked to disease.
Rna splicing
The document discusses RNA splicing and the spliceosome. It notes that Phillip Allen Sharp co-discovered RNA splicing and won the 1993 Nobel Prize for discovering that eukaryotic genes contain introns that are removed from messenger RNA through splicing. It then provides details about the spliceosome, a large molecular machine that helps remove introns. The spliceosome is made up of small nuclear ribonucleoproteins (snRNPs) including U1, U2, U4, U5 and U6, which act in complexes E, A, B and C during the splicing process.
basic techniques of biotechnology
The slides tells about the basic techniques performed in biotechnology lab. a initiator should be known with these techniques so that it become easier for the one who wants to see himself in a biotechnology field.
Nucleosome and chromatin structure
DNA is highly compacted in cells through various levels of supercoiling and chromatin structure. At the most basic level, DNA wraps around histone proteins to form nucleosomes, introducing negative supercoiling. Nucleosomes are then packed into a beads-on-a-string structure and further compacted into the 30nm fiber. Additional folding compacts the DNA over 10,000-fold into the final chromosomes. The two main types of supercoiling that facilitate compaction are plectonemic and solenoidal, with solenoidal supercoiling allowing the greatest degree of compaction and found in chromatin.
RNA Splicing
RNA splicing is a process in which introns are removed from pre-mRNA transcripts and exons are joined together to produce mature mRNA. There are three main types of splicing pathways: spliceosomal splicing, self-splicing, and tRNA splicing. Spliceosomal splicing involves the spliceosome complex and is the most common in eukaryotes. Self-splicing occurs without proteins through ribozyme activity. tRNA splicing uses ribonucleases and ligases. Alternative splicing allows different mRNA isoforms to be produced from the same pre-mRNA. Splicing errors can cause genetic diseases by disrupting protein sequences.
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Southern blotting
Southern Blotting is a technique developed by E.M. SOUTHERN BLOTTING in 1975. It includes Principle, steps and applications of southern blotting.
VBC-321_BLOTTING_TECHNIQUES and gel electrophoresis
Southern blotting is a technique used to detect specific DNA sequences. It involves separating DNA fragments by size via gel electrophoresis, transferring them to a membrane, then using a labeled probe to hybridize to complementary DNA sequences. This allows visualization of the location and size of DNA fragments containing the probe sequence. Key steps are digesting DNA, electrophoretic sorting, denaturing and transferring DNA to a membrane, blocking with excess DNA, washing unbound probe, then autoradiography to detect the bound probe and identify DNA fragments of interest.
Souther and northern blotting systems
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.
Blotting techniques
blotting techniques, western blotting, eastern blotting, northern blotting, southern blotting, immunoblotting, Far-eastern blotting, far western blotting
Blotting techniques.pptx
Blotting techniques involve the separation (via electrophoresis) and transfer of DNA, RNA, or proteins onto a blotting membrane. This separation is generally followed by complexing of the target with a labeled molecule for detection.
Used extensively in analysis of:
DNA.
RNA.
Proteins.
southern blotting.pptx
Blotting
A blot, in molecular biology and genetics, is a method of transferring proteins, DNA or RNA, onto a carrier.
The term "blotting" refers to the transfer of biological samples from a gel to a membrane and their subsequent detection on the surface of the membrane.
Types of blotting techniques
Southern Blotting
Northern Blotting
Western Blotting
A Southern blot is a method used
in molecular biology for detection of a specific DNA sequence in DNA samples.
Southern blotting combines transfer of electrophoresis -separated DNA fragments to a filter membrane and subsequent fragment detection by probe hybridization.
The method is named after its inventor, the British biologist Edwin Mellor Southern.
- Methods in Southern blotting
- Advantages and disadvantages
Southern-blotting-and-Western-blotting.pptx
1. DNA or proteins are separated by gel electrophoresis.
2. The molecules are then transferred to a membrane through blotting.
3. For Southern blotting, DNA is detected using labeled probes that hybridize to complementary DNA sequences. For Western blotting, proteins are detected using primary and secondary antibodies that bind to the protein of interest.
4. These techniques are used for applications like identifying genes, detecting infectious diseases and genetic disorders, and forensic analysis.
Southern blotting
Southern blotting and Northern blotting are techniques used to detect specific DNA and RNA sequences. Southern blotting involves extracting DNA from cells, cutting the DNA into fragments using restriction enzymes, separating the fragments by size using gel electrophoresis, transferring the fragments to a membrane, and using a labeled probe to detect complementary DNA sequences via hybridization. Northern blotting uses a similar process to detect specific RNA sequences, and is used to study gene expression and RNA processing. Both techniques allow detection of specific sequences among a large mixture of nucleic acids.
Southern blotting-1.pptx
Southern blotting is a technique used to detect specific DNA sequences. It involves extracting DNA, restricting it with enzymes, separating fragments via electrophoresis, transferring DNA to a membrane, then using a probe with a complementary sequence and radioactive tag to hybridize and identify the target DNA fragment through autoradiography. It is useful for detecting gene copies, identifying mutations, and diagnosing genetic diseases.
Blotting techniques includes southren,northern,western and dot blotting
This document describes various blotting techniques used to detect specific DNA or RNA sequences, including Southern blotting, Northern blotting, Western blotting, and dot blotting. Southern blotting involves transferring DNA fragments separated by gel electrophoresis to a membrane and probing for specific sequences. Northern blotting is similar but uses RNA. Western blotting detects specific proteins. Dot blotting detects sequences in non-fractionated samples by directly applying samples to a membrane. These techniques allow for detection and analysis of genetic material.
Blotting techniques
This document describes various blotting techniques used to detect specific DNA or RNA sequences, including Southern blotting, Northern blotting, Western blotting, and dot blotting. Southern blotting involves transferring DNA fragments separated by gel electrophoresis to a membrane and probing for specific sequences. Northern blotting is similar but uses RNA. Western blotting detects specific proteins. Dot blotting detects sequences in non-fractionated samples by directly applying samples to a membrane. These techniques allow for detection and analysis of genes and their expression.
blotting tech.pptx
The document discusses various techniques for DNA hybridization including Southern, Northern, Western, dot, and colony hybridization. Southern hybridization involves immobilizing DNA on a membrane, adding a labeled probe, and detecting hybridization to identify DNA sequences. Factors like stringency affect specificity. Northern hybridization identifies RNA using DNA probes. Western blotting identifies proteins using antibodies or lectins as probes. Dot and colony blotting modify Southern and Northern techniques to detect DNA or RNA from multiple samples immobilized on membranes.
Blotting technique including Southern , Northern and Western blotting
he given ppt contains all the blotting techniques which is being studied by students in Biotechnology related subject and this PPT contais all blotting techniques in a very elaborative concise manner includes procedure principle application etc so which itwould help any bio student to take proper knowledge in this topic. I hope you will enjoy the content of the topic and would be able to grasp the topic properly
VBC-321_BLOTTING_TECHNIQUES.pptx
1. Blotting techniques such as Southern, Northern, and Western blotting allow for the transfer of DNA, RNA, and proteins from a gel to a membrane for detection.
2. The Southern blot detects DNA using hybridization with a labeled probe. The Northern blot detects RNA and the Western blot detects proteins using antibodies.
3. These techniques separate biomolecules by size then transfer and detect them on a membrane using probes or antibodies, allowing analysis of complex samples.
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.
Presentation1
This presentation provides an overview of northern blotting. Northern blotting is a technique used to detect RNA in a sample. It involves separating RNA fragments via gel electrophoresis, transferring them to a membrane, then using a labeled probe to detect the RNA of interest. The key steps are isolating RNA from cells, separating fragments by size via gel electrophoresis, transferring RNA to a membrane, hybridizing the membrane with a labeled probe, and detecting the RNA of interest on the membrane. Northern blotting allows researchers to study gene expression patterns and RNA splicing.
blooting image.pptx
1. DNA or proteins are separated by gel electrophoresis.
2. The molecules are then transferred to a membrane through either southern or western blotting.
3. For southern blotting, DNA is detected through hybridization with a labeled probe. For western blotting, specific proteins are detected through interaction with labeled antibodies.
4. These techniques are used for diagnostic purposes and research applications such as identifying DNA sequences or proteins of interest.
Northern blotting
This presentation is all about Northern blotting technique ..which is a biotechnological process used in cell and molecular biology studies.
Molecular methods
Molecular methods are used to detect viruses that cannot be cultured in vitro by detecting their nucleic acids. Techniques like PCR and nucleic acid-based amplification are used to detect viral genomes. Other techniques include dot blot, Southern blot, Northern blot, and in situ hybridization. Newer techniques include PCR, ligase chain reaction, branched DNA, and nucleic acid-based amplification. Nucleic acid probes are short labeled DNA strands that anneal to complementary target DNA strands through base pairing. Southern blotting separates DNA fragments by size and transfers them to a filter for detection. Northern blotting separates mRNA and transfers it to a filter. Western blotting separates proteins by size and transfers them to a filter for immunodetection.
Similar to Southern blotting (20)
VBC-321_BLOTTING_TECHNIQUES and gel electrophoresis
VBC-321_BLOTTING_TECHNIQUES and gel electrophoresis
Blotting techniques includes southren,northern,western and dot blotting
Blotting techniques includes southren,northern,western and dot blotting
Blotting technique including Southern , Northern and Western blotting
Blotting technique including Southern , Northern and Western blotting
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Southern blotting
- 1. Southern Blotting BY SYEDA BATOOL ABBAS
- 2. What is Southern blotting? Purpose : It is used for DNA analysis for the detection of specific DNA sequences in a DNA sample. Also known as DNA blotting . Technique developed by Edwin Southern in 1975 in Edinburg university. Method named after its inventor. Specify the presence or absence of a particular nucleotide Used in genetically modified organisms.
- 3. Principle To Extract, purify and identify specific DNA sequences from other cells.
- 4. METHOD Step 1: DNA is separated from the source and cut into many small restriction fragments by digesting it with restriction enzymes.
- 5. METHOD Step 2 The complex mixture of fragments is subjected to gel electrophoresis to separate the fragments according to size. Shortest at the top longest at the bottom.
- 6. Step 3: Ethidium Bromide intercalates between the DNA strands making it visible. DNA is taken from the gel and placed in the nylon filter.
- 7. Step 4: A radioactive DNA probe is added to the complement ary DNA strands.
- 8. Step 5: Excess probe is washed away and the probe bound to the filter is detected by autoradiography, which reveals the DNA fragment to which the probe hybridized.
- 9. Overview
- 10. USES OF THE TECHNIQUE Southern blots are used in gene discovery and mapping, evolution and development studies, diagnostics and forensics. In regards to genetically modified organisms, Southern blotting is used as a definitive test to ensure that a particular section of DNA of known genetic sequence has been successfully incorporated into the genome of the host organism.
- 11. References 1) http://highered.mheducation.com/sites/0072556781/student_view0/chapter14/animation_ quiz_5.html 2) http://www.bio.davidson.edu/genomics/method/Southernblot.html 3) https://askabiologist.asu.edu/southern-blotting
- 12. ???
- 13. Thankyou !