Genomic in situ hybridization (GISH) is a cytogenetic technique that allows radiolabeling of genomic DNA within cells, allowing visualization under a fluorescence microscope. GISH was developed in 1986 for animal hybrid cell lines and 1987 for plant studies. It involves extracting and radiolabeling genomic DNA from one organism as a probe to target and hybridize to similar genomic regions of another organism. Unhybridized regions can then be stained, allowing visualization of probe-target complexes and unlabeled regions. GISH provides quick, sensitive, and informative results for establishing phylogenetic relationships and identifying hybridized genomes.
One of the first plausible models to account for the preceding observations was
formulated by Robin Holliday.
The key features of the Holliday model are the formation of heteroduplex DNA; the
creation of a cross bridge; its migration along the two heteroduplex strands,
termed branch migration; the occurrence of mismatch repair; and the
subsequent resolution, or splicing, of the intermediate structure to yield different
typesof recombinant molecules.
Basics of Undergraduate/university fellows
Nucleosome model of chromosome is proposed by ROGER KORNBERG (son of Arthur
Kornberg) in 1974.
It was confirmed and crystalised by P. Oudet et al., (1975).
Nucleosome is the lowest level of Chromosome organization in eukaryotic cells.
Nucleosome model is a scientific model which explains the organization of DNA and
associated proteins in the chromosomes.
Nucleosome model also explains the exact mechanism of the folding of DNA in
thenucleus.
It is the most accepted model of chromatin organization.
It is used to identify chromosomal rearrangements in cancer patients.
Chromosomal identification in cell.
Detect the specific nucleotide sequence within cell and tissues.
Unique point among the studies of cell, biology, cytogenetics and molecular genetics
It is possible to detect single copy sequence on chromosome with probes.
genomic in situ hybridization (GISH) is a potentially powerful tool for studying genome evolution and biosystematics
It will useful for investigating the origins of wild and cultivated polyploid plant species
RAPD markers are decamer DNA fragments.
RAPD is a type of PCR reaction.
as the name suggest it is a fast method when compared to the traditional PCR medthod.
Techniques based on the principle of selectively amplifying a subset of restriction fragments from a complex mixture of DNA fragments obtained after digestion of genomic DNA with restriction endonucleases.
Chromosomes are known as hereditary vehicles
They are formed of strands of DNA molecules which contain information for the development of different characteristics and performance of various metabolic activities of the cells
The coordination of various function is brought about through the formation of enzymes which are complex protein molecules
A physical map of a chromosome or a genome that shows the physical locations of genes and other DNA sequences of interest. Physical maps are used to help scientists identify and isolate genes by positional cloning.
According to the ICSM (Intergovernmental Committee on Surveying and Mapping), there are five different types of maps: General Reference, Topographical, Thematic, Navigation Charts and Cadastral Maps and Plans.
Surface Plasmon Resonance,
Surface Plasmons:
Plasmons confined to surface (interface) and interact with light resulting in polarities.
Propagating electron density waves occurring at the interface between metal and dielectric.
One of the first plausible models to account for the preceding observations was
formulated by Robin Holliday.
The key features of the Holliday model are the formation of heteroduplex DNA; the
creation of a cross bridge; its migration along the two heteroduplex strands,
termed branch migration; the occurrence of mismatch repair; and the
subsequent resolution, or splicing, of the intermediate structure to yield different
typesof recombinant molecules.
Basics of Undergraduate/university fellows
Nucleosome model of chromosome is proposed by ROGER KORNBERG (son of Arthur
Kornberg) in 1974.
It was confirmed and crystalised by P. Oudet et al., (1975).
Nucleosome is the lowest level of Chromosome organization in eukaryotic cells.
Nucleosome model is a scientific model which explains the organization of DNA and
associated proteins in the chromosomes.
Nucleosome model also explains the exact mechanism of the folding of DNA in
thenucleus.
It is the most accepted model of chromatin organization.
It is used to identify chromosomal rearrangements in cancer patients.
Chromosomal identification in cell.
Detect the specific nucleotide sequence within cell and tissues.
Unique point among the studies of cell, biology, cytogenetics and molecular genetics
It is possible to detect single copy sequence on chromosome with probes.
genomic in situ hybridization (GISH) is a potentially powerful tool for studying genome evolution and biosystematics
It will useful for investigating the origins of wild and cultivated polyploid plant species
RAPD markers are decamer DNA fragments.
RAPD is a type of PCR reaction.
as the name suggest it is a fast method when compared to the traditional PCR medthod.
Techniques based on the principle of selectively amplifying a subset of restriction fragments from a complex mixture of DNA fragments obtained after digestion of genomic DNA with restriction endonucleases.
Chromosomes are known as hereditary vehicles
They are formed of strands of DNA molecules which contain information for the development of different characteristics and performance of various metabolic activities of the cells
The coordination of various function is brought about through the formation of enzymes which are complex protein molecules
A physical map of a chromosome or a genome that shows the physical locations of genes and other DNA sequences of interest. Physical maps are used to help scientists identify and isolate genes by positional cloning.
According to the ICSM (Intergovernmental Committee on Surveying and Mapping), there are five different types of maps: General Reference, Topographical, Thematic, Navigation Charts and Cadastral Maps and Plans.
Surface Plasmon Resonance,
Surface Plasmons:
Plasmons confined to surface (interface) and interact with light resulting in polarities.
Propagating electron density waves occurring at the interface between metal and dielectric.
Genomic In-Situ Hybridization (GISH)-Principles, Methods and Applications in ...Banoth Madhu
Banoth Madhu: Genomic In-Situ Hybridization (GISH)-Principles, Methods and Applications in Crop Plants. It is a cytogenetic technique that allows the detection and localization of specific nucleic acid sequences on morphologically preserved chromosomes using genomic DNA of donor specie as probe. It is a cytogenetic technique that allows the detection and localization of specific nucleic acid sequences on morphologically preserved chromosomes using genomic DNA of donor specie as probe
Fluorescent in situ hybridization (FISH) is a cytogenetic technique that can be used to detect and localize the presence or absence of specific DNA sequences on chromosomes.
Advances in Molecular Cytogenetics: Potential for Crop Improvement.pptxKanshouwaModunshim
Title: Exploring Advances in Cytogenetics and Molecular Cytogenetics
Description:
Delve into the intricate world of cytogenetics and its cutting-edge counterpart, molecular cytogenetics, through this insightful presentation. Understand the profound relationship between chromosome structure, behavior, and gene function, with a particular focus on their relevance to crop improvement programs.
Key Points:
Introduction to Cytogenetics: Explore the fundamental principles of cytogenetics, its historical significance, and the recent influence of molecular tools, leading to the emergence of molecular cytogenetics.
Importance in Crop Improvement: Uncover the pivotal role of molecular cytogenetics in crop improvement programs, offering insights into the structural and functional organization of genomes within chromosomes.
Karyotyping: Gain a comprehensive understanding of karyotyping, its significance in identifying chromosomal abnormalities, and its applications in studying evolutionary relationships among different taxa.
Chromosome Identification and Sorting: Learn about the techniques involved in the identification and sorting of individual chromosomes, crucial steps in cytogenetics research for various crops.
Chromosome Banding Techniques: Explore different chromosome banding techniques, such as G-Banding and C-Banding, and understand their applications in detecting structural rearrangements.
CHIAS (Chromosome Image Analyzing System): Get insights into the CHIAS software and its role in mapping and identifying chromosomes automatically.
Flow Cytometry: Discover the applications of flow cytometry in detecting and measuring physical and chemical characteristics of cells, with a focus on its relevance in chromosome research.
In Situ Hybridization: Explore the technique of in situ hybridization, particularly the fluorescent variant, and its applications in precise localization of specific DNA segments.
Genomics and Whole Genome Sequencing: Delve into the realm of genomics and whole-genome sequencing, understanding the approaches like BAC to BAC and Whole Genome Shotgun.
Case Study: Uncover a case study involving the identification of a Wheat-Psathyrostachys huashanica ditelosomic addition line, showcasing the practical applications of the discussed techniques.
Conclusion: Summarize the key takeaways from the presentation, emphasizing the role of these techniques in advancing precision breeding and crop improvement.
Definition, principle, Chemical used during the process, application, advantages, and disadvantages of both techniques. along with relevant case study for better understand
Industrial Production of L-Lysine by FermentationKuldeep Sharma
Lysine is an essential amino acid that is used in the biosynthesis of proteins. Lysine is required for the nutrition of animals and humans. Lysine is useful as medicament, chemical agent, food material (food industry) and feed additives (animal food). It's demand has been steadily increasing in recent years. Several thousand tones of L-lysine are annually produced worldwide, almost by microbial fermentation.
±For Education Purpose Only
Patanjali Ayurved Limited is an Indian FMCG company. Manufacturing units and headquarters are located in the industrial area of Haridwar while the registered office is located at Delhi. The company manufactures mineral and herbal products.
±For Education Purpose Only
DNA Replication stress is any hindrance to progression or movement of the replication fork. Recently, its been related to genomic instability and cancer.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
3. •“Genomic in situ hybridization (GISH) is a cytogenetic technique
that allows one to radiolabel parts of genome within the cells.”
•GISH was mainly developed for the animal hybrid cell lines (1986)
and later used for the plants at Plant Breeding Institute, Cambridge
(1987), where this technique got its name.
•The GISH is quick, sensitive, accurate, informative and a
comparative approach rather than absolute one.
• GISH technique is an advancement in the fluorescence in situ
hybridization (FISH) technique.
4.
5. -
•The technique involves the extraction and sub-
sequentially radio labeling of whole DNA of one organism
and to use as a probe to target the genome of another
organism.
•The parts of genome that are sufficiently similar to the
probe hybridize to form a probe-target complex which is
now labeled.
•The remaining unhybridized parts of genome can be
further stained to view them.
8. The Main steps involved in the genomic in situ hybridization are :
1. Direct or indirect labeling of probe.
2. Blocking DNA fragmentation
3. Preparation of slide.
4. Denaturation of Probe and blocking DNA in a hybridization mixture.
5. Addition of the probe and the blocking DNA with the hybridization
mixture.
6. Chromosome DNA denaturation.
7. Hybridization of blocking DNA and probe in the target sequence of
the chromosome.
8. Detection of the probe in the chromosome of one parent.
9. Chromosome DNA molecule of the second parent related to the
unlabeled blocking DNA.
10. Visualization of hybridization signals in a fluorescence microscope.
Unlabelled chromosomes are visualized with a counter-stain (blue).
11. Figure: Observed under Fluorescence Microscope
Source: Brammer et al (2013)
Counter-stained (Blue) Labeled Merged Image (A+B)
12. Genomic in situ Hybridisation
• Radio labeling of genome
• Quick results
• Fluorescence microscope is
used
• Site Specific
3D Genome Sequencing
• Restriction cleavage of
genome
• Time consuming technique
• Hi-C matrix and models are
used
• Multiple step technique
13.
14. Genomic in situ Hybridisation
• Probe : Entire genome.
• Comparative Approach.
• Main application:
Establishing phylogenetic
relationship.
Fluorescence in situ Hybridization
• Probe: Oligonucleotide
sequence.
• Absolute Approach.
• Main Application:
Determination of
Repetitive DNA Sequence.
15.
16. The major application of GISH technique are as
follows:
1. Meiotic studies
2. Determination of phylogenetic relationship.
3. Determine the positions of translocation
breakpoints
4. Comparative genomic studies of malignant and
normal cells of an individual
5. Unknown genome identification
6. To identify the hybridized genome of crop varieties