Principle and applications of blotting techniquesJayeshRajput7
The document discusses various blotting techniques used in molecular biology including Northern blotting, Southern blotting, dot blotting, colony hybridization, and plaque hybridization.
Northern blotting involves separating RNA samples by size, transferring them to a membrane, and using a probe to detect specific sequences. Southern blotting is used to detect specific DNA sequences by separating DNA fragments, transferring them to a membrane, and using probes. Dot blotting simplifies the detection of proteins by applying samples directly to a membrane. Colony hybridization screens bacterial colonies for genes of interest by transferring DNA to a membrane and using probes. Plaque hybridization identifies recombinant phages using a similar process to colony hybridization.
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.
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.
Principle and application of blotting techniquesJayeshRajput7
This document discusses various blotting techniques used in molecular biology including Northern blotting, Southern blotting, dot blotting, colony hybridization, and plaque hybridization. Northern blotting is used to detect RNA, Southern blotting detects specific DNA sequences, dot blotting detects proteins without separation, colony hybridization screens bacterial colonies for desired genes, and plaque hybridization identifies recombinant phages. These techniques allow for detection and analysis of nucleic acids and proteins to study gene expression, mutations, genetic diseases, and more.
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.
The document discusses various techniques used for nucleic acid hybridization, including Southern blotting, Northern blotting, dot blot hybridization, and in situ hybridization. Southern blotting involves separating DNA fragments by size, transferring them to a membrane, and using a labeled probe to detect complementary DNA sequences. It can be used to detect mutations. Northern blotting is similar but detects RNA. Dot blot hybridization spots DNA/RNA samples directly onto a membrane. In situ hybridization detects nucleic acids within intact cells using labeled probes. Microarrays allow simultaneous screening of thousands of genes using hybridization on an array.
This document summarizes different methods for screening DNA libraries to identify specific clones. It discusses screening by hybridization using radioactive probes or alternative labeling methods. It also describes screening by PCR using gene-specific primers and screening expression libraries using antibodies that recognize antigenic determinants on expressed polypeptides. Screening methods like Southwestern and Northwestern blotting combine principles of Southern/Western blots to identify DNA or RNA binding proteins. The goal of these screening methods is to efficiently identify clones containing specific DNA sequences or expressing desired proteins from large DNA libraries.
Principle and applications of blotting techniquesJayeshRajput7
The document discusses various blotting techniques used in molecular biology including Northern blotting, Southern blotting, dot blotting, colony hybridization, and plaque hybridization.
Northern blotting involves separating RNA samples by size, transferring them to a membrane, and using a probe to detect specific sequences. Southern blotting is used to detect specific DNA sequences by separating DNA fragments, transferring them to a membrane, and using probes. Dot blotting simplifies the detection of proteins by applying samples directly to a membrane. Colony hybridization screens bacterial colonies for genes of interest by transferring DNA to a membrane and using probes. Plaque hybridization identifies recombinant phages using a similar process to colony hybridization.
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.
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.
Principle and application of blotting techniquesJayeshRajput7
This document discusses various blotting techniques used in molecular biology including Northern blotting, Southern blotting, dot blotting, colony hybridization, and plaque hybridization. Northern blotting is used to detect RNA, Southern blotting detects specific DNA sequences, dot blotting detects proteins without separation, colony hybridization screens bacterial colonies for desired genes, and plaque hybridization identifies recombinant phages. These techniques allow for detection and analysis of nucleic acids and proteins to study gene expression, mutations, genetic diseases, and more.
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.
The document discusses various techniques used for nucleic acid hybridization, including Southern blotting, Northern blotting, dot blot hybridization, and in situ hybridization. Southern blotting involves separating DNA fragments by size, transferring them to a membrane, and using a labeled probe to detect complementary DNA sequences. It can be used to detect mutations. Northern blotting is similar but detects RNA. Dot blot hybridization spots DNA/RNA samples directly onto a membrane. In situ hybridization detects nucleic acids within intact cells using labeled probes. Microarrays allow simultaneous screening of thousands of genes using hybridization on an array.
This document summarizes different methods for screening DNA libraries to identify specific clones. It discusses screening by hybridization using radioactive probes or alternative labeling methods. It also describes screening by PCR using gene-specific primers and screening expression libraries using antibodies that recognize antigenic determinants on expressed polypeptides. Screening methods like Southwestern and Northwestern blotting combine principles of Southern/Western blots to identify DNA or RNA binding proteins. The goal of these screening methods is to efficiently identify clones containing specific DNA sequences or expressing desired proteins from large DNA libraries.
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.
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.
This document discusses different types of DNA libraries and methods for screening libraries to identify clones containing genes of interest. It describes genomic and cDNA libraries, noting that genomic libraries contain all DNA fragments from an organism's genome while cDNA libraries contain only coding sequences. The key screening methods discussed are colony/plaque hybridization using radiolabeled probes, expression screening using antibodies, and PCR screening using gene-specific primers.
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.
The document discusses the process of synthesizing cDNA from mRNA. It involves isolating mRNA, using reverse transcriptase to copy the mRNA into single-stranded cDNA, then converting it to double-stranded cDNA using DNA polymerase. The double-stranded cDNA can then be inserted into a vector and used to create a cDNA library through cloning in bacteria or phage. The library can be screened by hybridization or assays to identify clones containing genes of interest.
1) The document describes a new DNA sequencing system that uses picolitre-sized reactors on a microfabricated chip to massively parallelize sequencing reactions and significantly increase throughput over previous methods.
2) Key aspects of the system include using emulsion PCR to amplify DNA fragments on beads in picolitre droplets and performing pyrosequencing reactions in the high-density array of picolitre wells on the chip.
3) The document demonstrates the capabilities of the system by sequencing the genome of Mycoplasma genitalium and assembling it de novo, achieving 96% coverage of the 580 kb genome in a single run at 99.96% accuracy.
Nucleic acid hybridization is a technique to detect specific nucleic acid sequences. It involves allowing single-stranded DNA or RNA molecules to interact and form hybrids based on complementary base pairing. This allows researchers to determine the degree of sequence identity between nucleic acids and detect specific sequences. Fluorescent in situ hybridization (FISH) is a type of hybridization that uses fluorescent probes to map DNA sequences onto chromosomes. FISH has applications in cytogenetics, cancer detection, and prenatal diagnosis of genetic disorders.
2017 summer high school textbook (molecular biology and neuroscience)Masuma Sani
The document provides details about various techniques that will be covered in the Neuroscience Camp for High School Students. The camp will include hands-on activities involving 1) DNA techniques like PCR and electrophoresis, 2) protein techniques like Western blotting, 3) electrophysiology to record ion channels, 4) techniques to deliver genes to the brain and examine brain tissue, and 5) behavioral tests to assess learning and memory. Students will gain experience with fundamental molecular and cellular neuroscience methods.
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.
Blotting techniques such as Southern, northern, and western blotting are used to identify unique proteins and nucleic acid sequences. Southern blotting detects DNA sequences, northern blotting detects RNA, and western blotting detects proteins. The general procedure involves separating molecules by electrophoresis, transferring them to a membrane, and using probes to detect the molecule of interest through autoradiography or another detection method. DNA microarrays can also detect gene expression levels but are currently too expensive for routine use.
1. DNA libraries contain cloned DNA fragments from an organism that can be used to isolate specific genes of interest. There are two main types: genomic libraries containing all DNA, and cDNA libraries containing expressed mRNA.
2. Bacterial transformation is a process where bacteria take up foreign DNA through horizontal gene transfer. It is used to clone and replicate plasmids in bacteria.
3. Chromosomal jumping and walking are techniques used in genome mapping and sequencing. Jumping allows bypassing repetitive sequences, while walking maps contiguous overlapping fragments to identify adjacent sequences.
1. Northern blotting is a technique used to detect specific RNA sequences in a sample. It involves separating RNA fragments by gel electrophoresis, transferring them to a membrane, then using a complementary DNA or RNA probe to identify the target sequence through hybridization.
2. Key steps include isolating RNA from cells, separating fragments by size using gel electrophoresis, blotting onto a membrane, incubating with a labeled probe, washing unbound probe, and detecting the bound probe to identify the target RNA sequence.
3. Northern blots are useful for determining gene expression levels in different tissues and cell types by detecting the presence and amount of specific mRNA transcripts. They allow investigation of how genes are regulated at the transcriptional level.
This document discusses various blotting techniques used to detect specific DNA, RNA, and protein molecules. It describes Southern blotting for detecting DNA, Northern blotting for detecting RNA, and Western blotting for detecting proteins. Southern blotting involves separating DNA fragments by gel electrophoresis, transferring them to a membrane, and using a labeled probe for detection. Northern blotting is similar but used for detecting specific RNA sequences. Western blotting uses SDS-PAGE gel electrophoresis to separate proteins, transfers them to a membrane, and detects them using primary and secondary antibodies. These techniques allow detection of specific biomolecules among many contaminants and have various applications in research and diagnostics.
Southern blotting is a technique developed by Professor Edwin Southern in 1975 to detect specific DNA sequences. It involves separating DNA fragments by size, transferring them to a membrane, then using a probe to detect the bound fragment. It is used for gene mapping, evolution studies, and DNA fingerprinting. Northern blotting detects specific RNA sequences and was developed in 1979. RNA is separated by size and transferred to a membrane, then a probe detects bound RNA fragments. Western blotting detects specific proteins and was developed in 1981. Proteins are separated by gel electrophoresis, transferred to a membrane, and a primary antibody binds the target protein, which is then detected by a secondary antibody.
Different blots are used to identify the presence of one specific target molecule (DNA, RNA or protein) in a complex mixture of related molecules. Blotting refers to the transfer of macromolecules (nucleic acids, proteins) from a gel onto the solid surface of an immobilized membrane for the detection of the transferred molecules.
Gene libraries, such as cDNA and genomic libraries, allow isolation of specific genes. cDNA libraries contain only exons and reflect gene expression levels, while genomic libraries contain all DNA fragments. Libraries are constructed by fragmenting DNA and cloning into vectors before transforming bacteria. They can be screened by hybridization, PCR, or immunological assays to detect gene products. Common steps include lysis, fixation, and detection to identify positive clones containing genes of interest.
Tracking introgressions using FISH and GISHvipulkelkar1
FISH and GISH are powerful cytogenetic techniques that allow the detection and localization of specific DNA sequences on chromosomes. FISH uses fluorescent probes to visualize DNA locations, while GISH uses total genomic DNA as probes. Both techniques have various applications, including chromosome mapping, analyzing hybrid plants and somatic variations, and detecting chromosomal abnormalities. They have improved plant breeding and furthered understanding of plant genomes, evolution, and relationships. Limitations include inability to detect small mutations and lack of commercial probes for all regions.
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.
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.
This document discusses different types of DNA libraries and methods for screening libraries to identify clones containing genes of interest. It describes genomic and cDNA libraries, noting that genomic libraries contain all DNA fragments from an organism's genome while cDNA libraries contain only coding sequences. The key screening methods discussed are colony/plaque hybridization using radiolabeled probes, expression screening using antibodies, and PCR screening using gene-specific primers.
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.
The document discusses the process of synthesizing cDNA from mRNA. It involves isolating mRNA, using reverse transcriptase to copy the mRNA into single-stranded cDNA, then converting it to double-stranded cDNA using DNA polymerase. The double-stranded cDNA can then be inserted into a vector and used to create a cDNA library through cloning in bacteria or phage. The library can be screened by hybridization or assays to identify clones containing genes of interest.
1) The document describes a new DNA sequencing system that uses picolitre-sized reactors on a microfabricated chip to massively parallelize sequencing reactions and significantly increase throughput over previous methods.
2) Key aspects of the system include using emulsion PCR to amplify DNA fragments on beads in picolitre droplets and performing pyrosequencing reactions in the high-density array of picolitre wells on the chip.
3) The document demonstrates the capabilities of the system by sequencing the genome of Mycoplasma genitalium and assembling it de novo, achieving 96% coverage of the 580 kb genome in a single run at 99.96% accuracy.
Nucleic acid hybridization is a technique to detect specific nucleic acid sequences. It involves allowing single-stranded DNA or RNA molecules to interact and form hybrids based on complementary base pairing. This allows researchers to determine the degree of sequence identity between nucleic acids and detect specific sequences. Fluorescent in situ hybridization (FISH) is a type of hybridization that uses fluorescent probes to map DNA sequences onto chromosomes. FISH has applications in cytogenetics, cancer detection, and prenatal diagnosis of genetic disorders.
2017 summer high school textbook (molecular biology and neuroscience)Masuma Sani
The document provides details about various techniques that will be covered in the Neuroscience Camp for High School Students. The camp will include hands-on activities involving 1) DNA techniques like PCR and electrophoresis, 2) protein techniques like Western blotting, 3) electrophysiology to record ion channels, 4) techniques to deliver genes to the brain and examine brain tissue, and 5) behavioral tests to assess learning and memory. Students will gain experience with fundamental molecular and cellular neuroscience methods.
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.
Blotting techniques such as Southern, northern, and western blotting are used to identify unique proteins and nucleic acid sequences. Southern blotting detects DNA sequences, northern blotting detects RNA, and western blotting detects proteins. The general procedure involves separating molecules by electrophoresis, transferring them to a membrane, and using probes to detect the molecule of interest through autoradiography or another detection method. DNA microarrays can also detect gene expression levels but are currently too expensive for routine use.
1. DNA libraries contain cloned DNA fragments from an organism that can be used to isolate specific genes of interest. There are two main types: genomic libraries containing all DNA, and cDNA libraries containing expressed mRNA.
2. Bacterial transformation is a process where bacteria take up foreign DNA through horizontal gene transfer. It is used to clone and replicate plasmids in bacteria.
3. Chromosomal jumping and walking are techniques used in genome mapping and sequencing. Jumping allows bypassing repetitive sequences, while walking maps contiguous overlapping fragments to identify adjacent sequences.
1. Northern blotting is a technique used to detect specific RNA sequences in a sample. It involves separating RNA fragments by gel electrophoresis, transferring them to a membrane, then using a complementary DNA or RNA probe to identify the target sequence through hybridization.
2. Key steps include isolating RNA from cells, separating fragments by size using gel electrophoresis, blotting onto a membrane, incubating with a labeled probe, washing unbound probe, and detecting the bound probe to identify the target RNA sequence.
3. Northern blots are useful for determining gene expression levels in different tissues and cell types by detecting the presence and amount of specific mRNA transcripts. They allow investigation of how genes are regulated at the transcriptional level.
This document discusses various blotting techniques used to detect specific DNA, RNA, and protein molecules. It describes Southern blotting for detecting DNA, Northern blotting for detecting RNA, and Western blotting for detecting proteins. Southern blotting involves separating DNA fragments by gel electrophoresis, transferring them to a membrane, and using a labeled probe for detection. Northern blotting is similar but used for detecting specific RNA sequences. Western blotting uses SDS-PAGE gel electrophoresis to separate proteins, transfers them to a membrane, and detects them using primary and secondary antibodies. These techniques allow detection of specific biomolecules among many contaminants and have various applications in research and diagnostics.
Southern blotting is a technique developed by Professor Edwin Southern in 1975 to detect specific DNA sequences. It involves separating DNA fragments by size, transferring them to a membrane, then using a probe to detect the bound fragment. It is used for gene mapping, evolution studies, and DNA fingerprinting. Northern blotting detects specific RNA sequences and was developed in 1979. RNA is separated by size and transferred to a membrane, then a probe detects bound RNA fragments. Western blotting detects specific proteins and was developed in 1981. Proteins are separated by gel electrophoresis, transferred to a membrane, and a primary antibody binds the target protein, which is then detected by a secondary antibody.
Different blots are used to identify the presence of one specific target molecule (DNA, RNA or protein) in a complex mixture of related molecules. Blotting refers to the transfer of macromolecules (nucleic acids, proteins) from a gel onto the solid surface of an immobilized membrane for the detection of the transferred molecules.
Gene libraries, such as cDNA and genomic libraries, allow isolation of specific genes. cDNA libraries contain only exons and reflect gene expression levels, while genomic libraries contain all DNA fragments. Libraries are constructed by fragmenting DNA and cloning into vectors before transforming bacteria. They can be screened by hybridization, PCR, or immunological assays to detect gene products. Common steps include lysis, fixation, and detection to identify positive clones containing genes of interest.
Tracking introgressions using FISH and GISHvipulkelkar1
FISH and GISH are powerful cytogenetic techniques that allow the detection and localization of specific DNA sequences on chromosomes. FISH uses fluorescent probes to visualize DNA locations, while GISH uses total genomic DNA as probes. Both techniques have various applications, including chromosome mapping, analyzing hybrid plants and somatic variations, and detecting chromosomal abnormalities. They have improved plant breeding and furthered understanding of plant genomes, evolution, and relationships. Limitations include inability to detect small mutations and lack of commercial probes for all regions.
ESR spectroscopy in liquid food and beverages.pptxPRIYANKA PATEL
With increasing population, people need to rely on packaged food stuffs. Packaging of food materials requires the preservation of food. There are various methods for the treatment of food to preserve them and irradiation treatment of food is one of them. It is the most common and the most harmless method for the food preservation as it does not alter the necessary micronutrients of food materials. Although irradiated food doesn’t cause any harm to the human health but still the quality assessment of food is required to provide consumers with necessary information about the food. ESR spectroscopy is the most sophisticated way to investigate the quality of the food and the free radicals induced during the processing of the food. ESR spin trapping technique is useful for the detection of highly unstable radicals in the food. The antioxidant capability of liquid food and beverages in mainly performed by spin trapping technique.
The binding of cosmological structures by massless topological defectsSérgio Sacani
Assuming spherical symmetry and weak field, it is shown that if one solves the Poisson equation or the Einstein field
equations sourced by a topological defect, i.e. a singularity of a very specific form, the result is a localized gravitational
field capable of driving flat rotation (i.e. Keplerian circular orbits at a constant speed for all radii) of test masses on a thin
spherical shell without any underlying mass. Moreover, a large-scale structure which exploits this solution by assembling
concentrically a number of such topological defects can establish a flat stellar or galactic rotation curve, and can also deflect
light in the same manner as an equipotential (isothermal) sphere. Thus, the need for dark matter or modified gravity theory is
mitigated, at least in part.
The cost of acquiring information by natural selectionCarl Bergstrom
This is a short talk that I gave at the Banff International Research Station workshop on Modeling and Theory in Population Biology. The idea is to try to understand how the burden of natural selection relates to the amount of information that selection puts into the genome.
It's based on the first part of this research paper:
The cost of information acquisition by natural selection
Ryan Seamus McGee, Olivia Kosterlitz, Artem Kaznatcheev, Benjamin Kerr, Carl T. Bergstrom
bioRxiv 2022.07.02.498577; doi: https://doi.org/10.1101/2022.07.02.498577
Immersive Learning That Works: Research Grounding and Paths ForwardLeonel Morgado
We will metaverse into the essence of immersive learning, into its three dimensions and conceptual models. This approach encompasses elements from teaching methodologies to social involvement, through organizational concerns and technologies. Challenging the perception of learning as knowledge transfer, we introduce a 'Uses, Practices & Strategies' model operationalized by the 'Immersive Learning Brain' and ‘Immersion Cube’ frameworks. This approach offers a comprehensive guide through the intricacies of immersive educational experiences and spotlighting research frontiers, along the immersion dimensions of system, narrative, and agency. Our discourse extends to stakeholders beyond the academic sphere, addressing the interests of technologists, instructional designers, and policymakers. We span various contexts, from formal education to organizational transformation to the new horizon of an AI-pervasive society. This keynote aims to unite the iLRN community in a collaborative journey towards a future where immersive learning research and practice coalesce, paving the way for innovative educational research and practice landscapes.
Travis Hills of MN is Making Clean Water Accessible to All Through High Flux ...Travis Hills MN
By harnessing the power of High Flux Vacuum Membrane Distillation, Travis Hills from MN envisions a future where clean and safe drinking water is accessible to all, regardless of geographical location or economic status.
ESA/ACT Science Coffee: Diego Blas - Gravitational wave detection with orbita...Advanced-Concepts-Team
Presentation in the Science Coffee of the Advanced Concepts Team of the European Space Agency on the 07.06.2024.
Speaker: Diego Blas (IFAE/ICREA)
Title: Gravitational wave detection with orbital motion of Moon and artificial
Abstract:
In this talk I will describe some recent ideas to find gravitational waves from supermassive black holes or of primordial origin by studying their secular effect on the orbital motion of the Moon or satellites that are laser ranged.
Current Ms word generated power point presentation covers major details about the micronuclei test. It's significance and assays to conduct it. It is used to detect the micronuclei formation inside the cells of nearly every multicellular organism. It's formation takes place during chromosomal sepration at metaphase.
EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...Sérgio Sacani
Context. With a mass exceeding several 104 M⊙ and a rich and dense population of massive stars, supermassive young star clusters
represent the most massive star-forming environment that is dominated by the feedback from massive stars and gravitational interactions
among stars.
Aims. In this paper we present the Extended Westerlund 1 and 2 Open Clusters Survey (EWOCS) project, which aims to investigate
the influence of the starburst environment on the formation of stars and planets, and on the evolution of both low and high mass stars.
The primary targets of this project are Westerlund 1 and 2, the closest supermassive star clusters to the Sun.
Methods. The project is based primarily on recent observations conducted with the Chandra and JWST observatories. Specifically,
the Chandra survey of Westerlund 1 consists of 36 new ACIS-I observations, nearly co-pointed, for a total exposure time of 1 Msec.
Additionally, we included 8 archival Chandra/ACIS-S observations. This paper presents the resulting catalog of X-ray sources within
and around Westerlund 1. Sources were detected by combining various existing methods, and photon extraction and source validation
were carried out using the ACIS-Extract software.
Results. The EWOCS X-ray catalog comprises 5963 validated sources out of the 9420 initially provided to ACIS-Extract, reaching a
photon flux threshold of approximately 2 × 10−8 photons cm−2
s
−1
. The X-ray sources exhibit a highly concentrated spatial distribution,
with 1075 sources located within the central 1 arcmin. We have successfully detected X-ray emissions from 126 out of the 166 known
massive stars of the cluster, and we have collected over 71 000 photons from the magnetar CXO J164710.20-455217.
When I was asked to give a companion lecture in support of ‘The Philosophy of Science’ (https://shorturl.at/4pUXz) I decided not to walk through the detail of the many methodologies in order of use. Instead, I chose to employ a long standing, and ongoing, scientific development as an exemplar. And so, I chose the ever evolving story of Thermodynamics as a scientific investigation at its best.
Conducted over a period of >200 years, Thermodynamics R&D, and application, benefitted from the highest levels of professionalism, collaboration, and technical thoroughness. New layers of application, methodology, and practice were made possible by the progressive advance of technology. In turn, this has seen measurement and modelling accuracy continually improved at a micro and macro level.
Perhaps most importantly, Thermodynamics rapidly became a primary tool in the advance of applied science/engineering/technology, spanning micro-tech, to aerospace and cosmology. I can think of no better a story to illustrate the breadth of scientific methodologies and applications at their best.
PPT on Direct Seeded Rice presented at the three-day 'Training and Validation Workshop on Modules of Climate Smart Agriculture (CSA) Technologies in South Asia' workshop on April 22, 2024.
The debris of the ‘last major merger’ is dynamically youngSérgio Sacani
The Milky Way’s (MW) inner stellar halo contains an [Fe/H]-rich component with highly eccentric orbits, often referred to as the
‘last major merger.’ Hypotheses for the origin of this component include Gaia-Sausage/Enceladus (GSE), where the progenitor
collided with the MW proto-disc 8–11 Gyr ago, and the Virgo Radial Merger (VRM), where the progenitor collided with the
MW disc within the last 3 Gyr. These two scenarios make different predictions about observable structure in local phase space,
because the morphology of debris depends on how long it has had to phase mix. The recently identified phase-space folds in Gaia
DR3 have positive caustic velocities, making them fundamentally different than the phase-mixed chevrons found in simulations
at late times. Roughly 20 per cent of the stars in the prograde local stellar halo are associated with the observed caustics. Based
on a simple phase-mixing model, the observed number of caustics are consistent with a merger that occurred 1–2 Gyr ago.
We also compare the observed phase-space distribution to FIRE-2 Latte simulations of GSE-like mergers, using a quantitative
measurement of phase mixing (2D causticality). The observed local phase-space distribution best matches the simulated data
1–2 Gyr after collision, and certainly not later than 3 Gyr. This is further evidence that the progenitor of the ‘last major merger’
did not collide with the MW proto-disc at early times, as is thought for the GSE, but instead collided with the MW disc within
the last few Gyr, consistent with the body of work surrounding the VRM.
The technology uses reclaimed CO₂ as the dyeing medium in a closed loop process. When pressurized, CO₂ becomes supercritical (SC-CO₂). In this state CO₂ has a very high solvent power, allowing the dye to dissolve easily.
SCREENING OF DNA LIBRARIES : hybridization, immunological method.
1. SCREENING AND PRESERVATION OF DNA LIBRARIES
Introduction
Library screening is the process of identification of the clones carrying the gene
of interest. Screening relies on a unique property of a clone in a library. The
DNA libraries consist of a collection of probably many thousand clones in the
form of either plaques or colonies on a plate. Screening of libraries can be done by
following approaches based on-
• Detecting a particular DNA sequence
• Gene expression.
Methods for screening based on detecting a DNA sequence
1. Screening by hybridization
Nucleic acid hybridization is the most commonly used method of
library screening first developed by Grunstein and Hogness in1975 to
detect DNA sequences in transformed colonies using radioactive RNA
probes.
It relies on the fact that a single-stranded DNA molecule, used as a probe
can hybridize to its complementary sequence and identify the specific
sequences.
This method is quick, can handle a very large number of clones and used
in the identification of cDNA clones which are not full-length (and
therefore cannot be expressed).
The commonly used methods of hybridization are,
a) Colony hybridization
b) Plaque hybridization.
2. Colony hybridization
Colony hybridization, also known as replica plating, allows the
screening of colonies plated at high density using radioactive DNA
probes. This method can be used to screen plasmid or cosmid based
libraries.
1) Preparation of Master plate: First, inoculate the bacterial cell
suspension on the solid agar medium to prepare the master plate. After the
inoculation, the number of bacterial colonies will develop with different
plasmids which refer as “Master or Reference plate”.
2) Formation of replicas over a nitrocellulose filter: Transfer the
bacterial cells from the master plate on to the membrane or filter by the
means of “Nitrocellulose filter”. Press the nitrocellulose filter paper over
the surface of the master plate. This compression of the filter membrane
will form replicas or copies of the bacterial cells as that of the master
plate.
3) Treatment of filter paper with SDS: Treat the nitrocellulose filter
paper with the detergent like SDS (Sodium dodecyl sulfate) to lyse the
bacterial cells.
4) Treatment of filter paper with alkali: Treat the filter paper with the
alkali like sodium hydroxide in order to separate the DNA into single
strands.
3. 5) Fixation of DNA onto the filter paper: To fix the DNA onto the
nitrocellulose filter paper, either bake the filter paper at 80 degrees
Celsius or expose it to the UV light.
6) Addition of radioactive probe: Hybridize the nitrocellulose filter
paper containing imprints of the plasmid DNA by the addition of
radioactive RNA probe. This radioactive RNA probe will code the
desired gene of sequence from the bacterial cells.
7) Washing and Autoradiography: Wash the filter paper to remove
unbound probe particles. After that, expose the nitrocellulose filter
paper to the X-ray film by the method refer as “Autoradiography”. The
colony which will appear after autoradiography will refer as
“Autoradiogram” which carry the genes of interest.
8) Identification of the desired gene: Compare the developed
autoradiogram with the master plate to identify the colonies
containing a gene of interest. The cells which contain the
desired gene can grow in the liquid medium and can further process
for the isolation of recombinant plasmid DNA
4. Plaque hybridization
Plaque hybridization, also known as Plaque lift, was developed by Benton and Davis in
1977 and employs a filter lift method applied to phage plaques. This procedure is
successfully applied to the isolation of recombinant phage by nucleic acid hybridization
and probably is the most widely applied method of library screening. The method of
screening library by plaque hybridization is described below-
The nitrocellulose filter is applied to the upper surface of agar plates, making a
direct contact between plaques and filter.
The plaques contain phage particles, as well as a considerable amount of
unpackaged recombinant DNA which bind to the filter.
The DNA is denatured, fixed to the filter, hybridized with radioactive probes and
assayed by autoradiography.
Advantages
• This method results in a ‘cleaner’ background and distinct signal (less background
probe hybridization) for λ plaque screening due to less DNA transfer from the
bacterial host to the nitrocellulose membrane while lifting plaques rather than
bacterial colonies.
• Multiple screens can be performed from the same plate as plaques can be lifted
several times.
• Screening can be performed at very high density by screening small plaques.
High-density screening has the advantage that a large number of
recombinantclones can be screened for the presence of sequences homologous to
the probe in a single experiment.
6. Screening methods based on gene expression
1. Immunological screening
This involves the use of antibodies that specifically recognize antigenic determinants
on the polypeptide. It does not rely upon any particular function of the expressed
foreign protein, but requires an antibody specific to the protein.
Earlier immunoscreening methods employed radio-labeled primary antibodies to
detect antibody binding to the nitrocellulose sheet (Figure 4-5.3.1(a).). It is now
superseded by antibody sandwiches resulting in highly amplified signals. The
secondary antibody recognizes the constant region of the primary antibody and is,
additionally, conjugated to an easily assayable enzyme (e.g. horseradish
peroxidase or alkaline phosphatase) which can be assayed using colorimetric change
or emission of light using X-ray film.
In this technique, the cells are grown as colonies on master plates and transferred
to a solid matrix.
These colonies are subjected to lysis releasing the proteins which bind to the
matrix.
These proteins are treated with a primary antibody which specifically binds to the
protein (acts as antigen), encoded by the target DNA. The unbound antibodies are
removed by washing.
A secondary antibody is added which specifically binds to the primary antibody
removing the unbound antibodies by washing.
The secondary antibody carries an enzyme label (e.g., horse radishperoxidase or
alkaline phosphatase) bound to it which converts colorless substrate to colored
product. The colonies with positive results (i.e. colored spots) are identified and
subcultured from the master plate.
7. Figure 4-5.3.1(a). Schematic process of immunological screening (a) a nitrocellulose disk is placed onto the surface of an agar
plate containing the phage library. Both agar plate and disk are marked so as to realign them later. (b) When the
nitrocellulose disk is lifted off again, proteins released from the bacteria by phage lysis bind to the disk. (c) These proteins bind
to specific antibody. (d) Plaques formed by bacteriophage that express the protein bound to the antibody will be detected by
emission of light. The positive clones can be identified by realignment.
(Adapted from Lodge J. 2007.Gene cloning: principles and applications. Taylor & Francis Group)
Figure 4-5.3.1(b). Schematic process of immunological screening using antibody sandwich.
8. The main difficulty with antibody-based screening is to raise a specific antibody for
each protein to be detected by injecting a foreign protein or peptide into an animal. This
is a lengthy and costly procedure and can only be carried out successfully with proteins
produced in reasonably large amounts.
4-5.3.2. Screening by functional complementation
Functional complementation is the process of compensating a missing function in a
mutant cell by a particular DNA sequence for restoring the wild-type phenotype. If the
mutant cells are non-viable, the cells carrying the clone of interest can be positively
selected and isolated. It is a very powerful method of expression cloning and also useful
for identification of genes from an organism having same role as that of defective gene in
another organism. The selection and identification of positive clones is based on either
the gain of function or a visible change in phenotype.
For example, the functional complementation in transgenic mice for the isolation of
Shaker-2 gene applied by Probst et al in1988 shown in Figure 4-5.3.2.
Figure 4-5.3.2. Functional complementation in transgenic mice for isolation of Shaker-2 gene.
(Adapted from Primrose SB, Twyman RM. 2006. Principles of gene manipulation and genomics.7th
ed. Blackwell Publishing.)
9. The Shaker-2 mutation is due to the defective gene associated with human deafness
disorder. The BAC clone from the wild type mice are prepared and injected into the eggs
of Shaker-2 mutants. The resulting mice are then screened for the presence of wild type
phenotype. Thus the BAC clone carrying the functional Shaker-2 gene is identified which
encodes a cytoskeletal myosin protein. This method can be used for screening human
genomic libraries to identify equivalent human gene.
Drawbacks
• Presence of an assayable mutation within the host cell that can be compensated by
the foreign gene expression which in most cases is not available. In addition,
foreign genes may not fully compensate the mutations.
Applications
• This method can be used for the isolation of higher-eukaryotic genes (e.g.
Drosophila topoisomerase II gene, a number of human RNA polymerase II
transcription factors) from an organism.
• It can also be possible in transgenic animals and plants to clone a specific gene
from its functional homologue.