Northern Blotting.pptx
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Northern blotting is a technique to detect specific RNA sequences. It involves separating RNA samples by size through agarose gel electrophoresis, blotting the RNA onto a membrane, then using a labeled probe to detect the RNA of interest through autoradiography or chemiluminescence. Northern blotting is used to study gene expression at the mRNA level, detect transcript size and splicing, and confirm transgenic and knockout animal models. While it provides information about RNA expression patterns, it is less sensitive than newer techniques and quality can be affected by RNA degradation.
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Northern blotting
Northern blotting is a technique used to detect specific RNA sequences. It involves separating RNA samples by size using gel electrophoresis, transferring the RNA to a membrane, then using a labeled probe to detect RNA sequences of interest through hybridization and detection of the probe. Northern blotting allows researchers to compare expression patterns of an RNA sequence among different tissue samples or developmental stages. It is commonly used to study gene expression and RNA degradation.
Blotting techniques1
Concept: reannealing nucleic acids to identify sequence of interest.
Separates DNA/RNA in an agarose gel, then detects specific bands using probe and hybridization.
Hybridization takes advantage of the ability of a single stranded DNA or RNA molecule to find its complement, even in the presence of large amounts of unrelated DNA.
Allows detection of specific bands (DNA fragments or RNA molecules) that have complementary sequence to the probe.
Size bands and quantify abundance of molecule.
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.
Blotting techniques
This document summarizes several blotting techniques used in molecular biology to detect biomolecules like DNA, RNA, and proteins. Southern blotting detects specific DNA sequences, Northern blotting detects RNA, and Western blotting detects proteins. Eastern blotting detects post-translational modifications to proteins. Dot blotting directly applies biomolecule samples to a membrane as dots to detect presence or absence without separation by size. These techniques involve transferring molecules to a membrane, probing with a labeled probe, washing, and detecting hybridized probes.
Northern blotting
This presentation is all about Northern blotting technique ..which is a biotechnological process used in cell and molecular biology studies.
Nothern blotting
This document discusses northern blotting, which is a laboratory technique used to detect specific RNA molecules. It involves separating RNA fragments by gel electrophoresis, transferring them to a membrane, then using a labeled probe to detect the RNA of interest through hybridization and visualization. Key steps include RNA isolation, gel electrophoresis, transfer to membrane, hybridization with probes, washing, and detection of labeled probes. The technique allows detection of RNA size and splice variants, and membranes can be stored for future 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.
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.
Recommended
Northern blotting
Northern blotting is a technique used to detect specific RNA sequences. It involves separating RNA samples by size using gel electrophoresis, transferring the RNA to a membrane, then using a labeled probe to detect RNA sequences of interest through hybridization and detection of the probe. Northern blotting allows researchers to compare expression patterns of an RNA sequence among different tissue samples or developmental stages. It is commonly used to study gene expression and RNA degradation.
Blotting techniques1
Concept: reannealing nucleic acids to identify sequence of interest.
Separates DNA/RNA in an agarose gel, then detects specific bands using probe and hybridization.
Hybridization takes advantage of the ability of a single stranded DNA or RNA molecule to find its complement, even in the presence of large amounts of unrelated DNA.
Allows detection of specific bands (DNA fragments or RNA molecules) that have complementary sequence to the probe.
Size bands and quantify abundance of molecule.
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.
Blotting techniques
This document summarizes several blotting techniques used in molecular biology to detect biomolecules like DNA, RNA, and proteins. Southern blotting detects specific DNA sequences, Northern blotting detects RNA, and Western blotting detects proteins. Eastern blotting detects post-translational modifications to proteins. Dot blotting directly applies biomolecule samples to a membrane as dots to detect presence or absence without separation by size. These techniques involve transferring molecules to a membrane, probing with a labeled probe, washing, and detecting hybridized probes.
Northern blotting
This presentation is all about Northern blotting technique ..which is a biotechnological process used in cell and molecular biology studies.
Nothern blotting
This document discusses northern blotting, which is a laboratory technique used to detect specific RNA molecules. It involves separating RNA fragments by gel electrophoresis, transferring them to a membrane, then using a labeled probe to detect the RNA of interest through hybridization and visualization. Key steps include RNA isolation, gel electrophoresis, transfer to membrane, hybridization with probes, washing, and detection of labeled probes. The technique allows detection of RNA size and splice variants, and membranes can be stored for future 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.
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 4 northern and western blotting 2
Northern blotting is a technique used to detect specific RNA sequences. It involves separating RNA samples by electrophoresis in a gel, transferring the RNA to a membrane, and using a labeled probe to detect the RNA of interest. Western blotting is similar but detects specific proteins using antibodies as probes. It involves separating proteins by electrophoresis, transferring them to a membrane, and using primary and secondary antibodies to detect the protein of interest. Both techniques are used to study gene and protein expression.
Blotting
Blotting techniques such as Southern, Northern, and Western blotting allow for the detection and analysis of 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 via gel electrophoresis, transferring them to a membrane, then using a labeled probe for detection. Northern blotting detects RNA and was developed in 1979, using a similar process to Southern blotting but separating and detecting RNA. Western blotting detects proteins and was developed in 1981, using antibodies specific to the target protein for detection. These techniques are widely used in research and diagnostics.
Blotting Techniques
This document discusses various blotting techniques used to detect and analyze biomolecules like DNA, RNA, and proteins. It describes the Southern blot technique developed by Edwin Southern used to detect specific DNA sequences. It also discusses the Northern blot technique used to detect RNA, developed by James Alwine and George Stark. Finally, it summarizes the Western blot technique used to detect specific proteins by using antibodies, developed in 1981. These blotting techniques allow separation and detection of biomolecules through transfer and hybridization/binding reactions.
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.
Blotting techniques
Southern, Northern, and Western blotting techniques allow researchers to detect specific DNA, RNA, and protein sequences, respectively. Southern blotting involves separating DNA fragments via gel electrophoresis, transferring them to a membrane, and using a probe to identify specific sequences. Northern blotting is similar but detects RNA, and Western blotting detects proteins using antibodies. These techniques are used for applications like gene mapping, diagnostics, studying gene expression, and confirming transgenic organisms.
Dna finger printing
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...
BLOTTING TECHNIQUES FOR RNA, DNA AND PROTEINS.pptx
These slides explain the three types of blotting techniques and the macromolecules they detect.
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.
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.
NORTHERN BLOTTING.pptx
This document describes the process of northern blotting. Northern blotting is used to detect specific RNA molecules and analyze gene expression. It involves separating RNA samples by size using 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 tissue or cells, separating the RNA by size on a gel, blotting the RNA onto a membrane, hybridizing the membrane with a labeled probe, washing off excess probe, and visualizing where the probe bound to detect the RNA of interest. Northern blotting allows researchers to study gene expression patterns in different tissues, developmental stages, or disease states.
Genetic fingerprinting
Genetic fingerprinting is a technique used to distinguish individuals using DNA samples. It was invented in 1985 by Alec Jeffreys at the University of Leicester. The process involves extracting DNA from samples like blood or tissue, cutting the DNA into fragments using restriction enzymes, separating the fragments by size using gel electrophoresis, and analyzing patterns of bands to identify individuals. Newer short tandem repeat analysis examines repetitive DNA sequences that show great variability among people, allowing for highly accurate identification with probabilities of a match being as low as 1 in 60,000. Genetic fingerprinting has applications in forensic science, determining family relationships, and healthcare.
blottingtechniques.pptx different types of blotting technique
Blotting techniques such as Western blotting, Southern blotting, and Northern blotting allow researchers to detect specific proteins, DNA sequences, or RNA sequences within a sample. These techniques involve separating biomolecules using gel electrophoresis, transferring them to a membrane, and using a probe to identify the target molecule. Southern blotting specifically detects DNA, Northern blotting detects RNA, and Western blotting detects proteins. These techniques are widely used for applications like gene mapping, diagnostics, studying gene expression, and detecting genetically modified organisms.
blotting techniques southern northern Western
Blotting techniques such as Western blotting, Southern blotting, and Northern blotting allow researchers to detect specific proteins, DNA sequences, or RNA sequences within a sample. These techniques involve separating biomolecules via gel electrophoresis, transferring them to a membrane, then using a probe to identify the target molecule. Southern blotting specifically detects DNA, Northern blotting detects RNA, and Western blotting detects proteins. These techniques are widely used for applications like gene mapping, diagnostics, studying gene expression, and detecting genetically modified organisms.
Presentation dna fingerprinting
DNA profiling process, RFLP analysis, STR analysis by PCR, basic principle of dna fingerprinting, advantages and disadvantages of RFLP and STR analysis
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.
Blotting techniques
most effective slide for the medical student ,though you will find most of the things in short time..
Blotting techniques
Blotting techniques such as Southern blotting, Northern blotting, and Western blotting allow researchers to detect specific DNA, RNA, or protein molecules within complex biological samples. Southern blotting is used to detect DNA and involves separating DNA fragments by gel electrophoresis, transferring them to a membrane, then using a DNA probe to hybridize to the target sequence. Northern blotting detects RNA and uses a similar process but with RNA samples. Western blotting identifies specific proteins using gel electrophoresis, transfer to a membrane, and antibody probes against the target protein. These techniques are important tools for research in areas like gene discovery, molecular diagnostics, and studying gene expression.
Dna fingerprinting
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.
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.
Creation or Update of a Mandatory Field is Not Set in Odoo 17
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Lecture 4 northern and western blotting 2
Northern blotting is a technique used to detect specific RNA sequences. It involves separating RNA samples by electrophoresis in a gel, transferring the RNA to a membrane, and using a labeled probe to detect the RNA of interest. Western blotting is similar but detects specific proteins using antibodies as probes. It involves separating proteins by electrophoresis, transferring them to a membrane, and using primary and secondary antibodies to detect the protein of interest. Both techniques are used to study gene and protein expression.
Blotting
Blotting techniques such as Southern, Northern, and Western blotting allow for the detection and analysis of 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 via gel electrophoresis, transferring them to a membrane, then using a labeled probe for detection. Northern blotting detects RNA and was developed in 1979, using a similar process to Southern blotting but separating and detecting RNA. Western blotting detects proteins and was developed in 1981, using antibodies specific to the target protein for detection. These techniques are widely used in research and diagnostics.
Blotting Techniques
This document discusses various blotting techniques used to detect and analyze biomolecules like DNA, RNA, and proteins. It describes the Southern blot technique developed by Edwin Southern used to detect specific DNA sequences. It also discusses the Northern blot technique used to detect RNA, developed by James Alwine and George Stark. Finally, it summarizes the Western blot technique used to detect specific proteins by using antibodies, developed in 1981. These blotting techniques allow separation and detection of biomolecules through transfer and hybridization/binding reactions.
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.
Blotting techniques
Southern, Northern, and Western blotting techniques allow researchers to detect specific DNA, RNA, and protein sequences, respectively. Southern blotting involves separating DNA fragments via gel electrophoresis, transferring them to a membrane, and using a probe to identify specific sequences. Northern blotting is similar but detects RNA, and Western blotting detects proteins using antibodies. These techniques are used for applications like gene mapping, diagnostics, studying gene expression, and confirming transgenic organisms.
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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...
BLOTTING TECHNIQUES FOR RNA, DNA AND PROTEINS.pptx
These slides explain the three types of blotting techniques and the macromolecules they detect.
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.
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.
NORTHERN BLOTTING.pptx
This document describes the process of northern blotting. Northern blotting is used to detect specific RNA molecules and analyze gene expression. It involves separating RNA samples by size using 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 tissue or cells, separating the RNA by size on a gel, blotting the RNA onto a membrane, hybridizing the membrane with a labeled probe, washing off excess probe, and visualizing where the probe bound to detect the RNA of interest. Northern blotting allows researchers to study gene expression patterns in different tissues, developmental stages, or disease states.
Genetic fingerprinting
Genetic fingerprinting is a technique used to distinguish individuals using DNA samples. It was invented in 1985 by Alec Jeffreys at the University of Leicester. The process involves extracting DNA from samples like blood or tissue, cutting the DNA into fragments using restriction enzymes, separating the fragments by size using gel electrophoresis, and analyzing patterns of bands to identify individuals. Newer short tandem repeat analysis examines repetitive DNA sequences that show great variability among people, allowing for highly accurate identification with probabilities of a match being as low as 1 in 60,000. Genetic fingerprinting has applications in forensic science, determining family relationships, and healthcare.
blottingtechniques.pptx different types of blotting technique
Blotting techniques such as Western blotting, Southern blotting, and Northern blotting allow researchers to detect specific proteins, DNA sequences, or RNA sequences within a sample. These techniques involve separating biomolecules using gel electrophoresis, transferring them to a membrane, and using a probe to identify the target molecule. Southern blotting specifically detects DNA, Northern blotting detects RNA, and Western blotting detects proteins. These techniques are widely used for applications like gene mapping, diagnostics, studying gene expression, and detecting genetically modified organisms.
blotting techniques southern northern Western
Blotting techniques such as Western blotting, Southern blotting, and Northern blotting allow researchers to detect specific proteins, DNA sequences, or RNA sequences within a sample. These techniques involve separating biomolecules via gel electrophoresis, transferring them to a membrane, then using a probe to identify the target molecule. Southern blotting specifically detects DNA, Northern blotting detects RNA, and Western blotting detects proteins. These techniques are widely used for applications like gene mapping, diagnostics, studying gene expression, and detecting genetically modified organisms.
Presentation dna fingerprinting
DNA profiling process, RFLP analysis, STR analysis by PCR, basic principle of dna fingerprinting, advantages and disadvantages of RFLP and STR analysis
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.
Blotting techniques
most effective slide for the medical student ,though you will find most of the things in short time..
Blotting techniques
Blotting techniques such as Southern blotting, Northern blotting, and Western blotting allow researchers to detect specific DNA, RNA, or protein molecules within complex biological samples. Southern blotting is used to detect DNA and involves separating DNA fragments by gel electrophoresis, transferring them to a membrane, then using a DNA probe to hybridize to the target sequence. Northern blotting detects RNA and uses a similar process but with RNA samples. Western blotting identifies specific proteins using gel electrophoresis, transfer to a membrane, and antibody probes against the target protein. These techniques are important tools for research in areas like gene discovery, molecular diagnostics, and studying gene expression.
Dna fingerprinting
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.
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.
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Northern Blotting.pptx
- 1. Welcome to My Todays Presentation Topic: Northern Blotting Presented by: Sita Tappo
- 2. Northern Blotting Northern blotting is a technique for detection of specific RNA sequences. Northern blotting was developed by James Alwine and George Stark at Stanford University (1979) and was named such by analogy to Southern blotting
- 3. Steps involved in Northern blotting 1. RNA is isolated from several biological samples (e.g. various tissues, various developmental stages of same tissue etc.) * RNA is more susceptible to degradation than DNA.
- 4. Cont…… 2. Sample’s are loaded on gel and the RNA samples are separated according to their size on an agarose gel . • The resulting gel following after the electrophoresis run.
- 5. Cont…… 3. The gel is then blotted on a nylon membrane or a nitrocellulose filter paper/ diazobenzyloxymethyl papers by creating the sandwich arrangement.
- 6. Cont…… 4. The membrane is placed in a dish containing hybridization buffer with a labeled probe. • Thus, it will hybridize to the RNA on the blot that corresponds to the sequence of interest. 5. The membrane is washed to remove unbound probe.
- 7. Cont…… 6. The labeled probe is detected via autoradiography or via a chemiluminescence reaction (if a chemically labeled probe is used). In both cases this results in the formation of a dark band on an X-ray film. • Now the expression patterns of the sequence of interest in the different samples can be compared.
- 8. APPLICATIONS • A standard for the study of gene expression at the level of mRNA(messenger RNA transcripts) • Detection of mRNA transcript size • Study RNA degradation • Study RNA splicing • Study RNA half-life • Often used to confirm and check transgenic / knockout mice (animals)
- 9. Disadvantage of Northern plotting 1.The standard northern blot method is relatively less sensitive 2. Detection with multiple probes is a problem 3. If RNA samples are even slightly degraded by RNases, the quality of the data and quantitation of expression is quite negatively affected.
- 10. Thank You for Listening With Great Patience