In situ Hybridization (ISH) and Fluorescence in Situ Hybridization (FISH) Creative-Diagnostics
In situ Hybridization (ISH) and Fluorescence in Situ Hybridization (FISH) by Creative Diagnostics, learn more http://www.creative-diagnostics.com/in-situ-hybridization-and-fluorescence-in-situ-hybridization.htm
Genotyping by Sequencing is a robust,fast and cheap approach for high throughput marker discovery.It has applications in crop improvement programs by enhancing identification of superior genotypes.
Fluorescent in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes that bind to only those parts of the chromosome with a high degree of sequence complementarity. Creative Bioarray provides comprehensive FISH services and products to our clients.
A real-time polymerase chain reaction is a laboratory technique of molecular biology based on the polymerase chain reaction (PCR). It monitors the amplification of a targeted DNA molecule during the PCR, i.e. in real-time, and not at its end, as in conventional PCR.
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In situ Hybridization (ISH) and Fluorescence in Situ Hybridization (FISH) Creative-Diagnostics
In situ Hybridization (ISH) and Fluorescence in Situ Hybridization (FISH) by Creative Diagnostics, learn more http://www.creative-diagnostics.com/in-situ-hybridization-and-fluorescence-in-situ-hybridization.htm
Genotyping by Sequencing is a robust,fast and cheap approach for high throughput marker discovery.It has applications in crop improvement programs by enhancing identification of superior genotypes.
Fluorescent in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes that bind to only those parts of the chromosome with a high degree of sequence complementarity. Creative Bioarray provides comprehensive FISH services and products to our clients.
A real-time polymerase chain reaction is a laboratory technique of molecular biology based on the polymerase chain reaction (PCR). It monitors the amplification of a targeted DNA molecule during the PCR, i.e. in real-time, and not at its end, as in conventional PCR.
https://www.patreon.com/biotechlive
SUPPORT EDUCATION... SUPPORT US
"In field molecular diagnostics as an aid to disease management"EMPHASIS PROJECT
Insights about isothermal Polymerase Chain Reaction (PCR) assays and how they can be used to diagnose the presence of latent diseases in the field, including those which are especially difficult to identify. They will show how assays are developed, and how they may be used to improve disease management choices.
The target audience are researchers, agri-business and forestry experts, farmers and foresters and any other interested in plant health.
Do not hesitate to contact EMPHASIS project through Facebook, Twitter, email (emphasisproject@gmail.com) or through their website (http://www.emphasisproject.eu/) if you want to be updated on webinars dates and content and book a ticket.
To watch on Youtube: https://youtu.be/yFEG9uTEhdc
Fluorescent in situ hybridization (FISH) is a cytogenetic technique that uses fluorescent probes to investigate the presence of small, submicroscopic chromosomal changes that are beyond the resolution of karyotype analysis.
This PowerPoint presentation explain the concept,process and application of Fluorescence insitu hybridization.
Fundamentals of Fluorescence in situ Hybridization Amartya Pradhan
This presentation provides an insight into the fundamentals of in situ hybridization (ISH), especially fluorescence in situ hybridization. It is ideal for classroom lecture.
Deciphering DNA sequences is essential for virtually all branches of biological research. With the
advent of capillary electrophoresis (CE)-based Sanger sequencing, scientists gained the ability to
elucidate genetic information from any given biological system. This technology has become widely
adopted in laboratories around the world, yet has always been hampered by inherent limitations in
throughput, scalability, speed, and resolution that often preclude scientists from obtaining the essential
information they need for their course of study. To overcome these barriers, an entirely new technology
was required—Next-Generation Sequencing (NGS), a fundamentally different approach to sequencing
that triggered numerous ground-breaking discoveries and ignited a revolution in genomic science.
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.
Gene mapping | Genetic map | Physical Map | DNA Data Analysis (upgraded)NARC, Islamabad
Genes are useful markers but not ideal.
Mapped feature that are not genes are called DNA markers.
DNA markers must have at least two alleles to be useful.
DNA sequence features that satisfy this requirement are-
– Restriction Fragment Length Polymorphism (RFLP)
Southern hybridization
PCR
– Simple Sequence Length Polymorphism (SSLP)
– Single Nucleotide Polymorphism (SNP)
Mapping- determining the location of elements with in a genome, with respect to identifiable land marks.
Gene mapping describes the methods used to identify the locus of a gene and the distances between genes.
In simple mapping of genes to specific locations on chromosomes.
Two types
Genetic map
Physical Map
They are useful in predicting results of dihybrid and trihybrid crosses.
It allows geneticists to understand the overall complexity and genetic organization of a particular species.
Identify genes responsible for diseases.
Identify genes responsible for traits.
genetic maps are useful from an evolutionary point of view.
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.
"In field molecular diagnostics as an aid to disease management"EMPHASIS PROJECT
Insights about isothermal Polymerase Chain Reaction (PCR) assays and how they can be used to diagnose the presence of latent diseases in the field, including those which are especially difficult to identify. They will show how assays are developed, and how they may be used to improve disease management choices.
The target audience are researchers, agri-business and forestry experts, farmers and foresters and any other interested in plant health.
Do not hesitate to contact EMPHASIS project through Facebook, Twitter, email (emphasisproject@gmail.com) or through their website (http://www.emphasisproject.eu/) if you want to be updated on webinars dates and content and book a ticket.
To watch on Youtube: https://youtu.be/yFEG9uTEhdc
Fluorescent in situ hybridization (FISH) is a cytogenetic technique that uses fluorescent probes to investigate the presence of small, submicroscopic chromosomal changes that are beyond the resolution of karyotype analysis.
This PowerPoint presentation explain the concept,process and application of Fluorescence insitu hybridization.
Fundamentals of Fluorescence in situ Hybridization Amartya Pradhan
This presentation provides an insight into the fundamentals of in situ hybridization (ISH), especially fluorescence in situ hybridization. It is ideal for classroom lecture.
Deciphering DNA sequences is essential for virtually all branches of biological research. With the
advent of capillary electrophoresis (CE)-based Sanger sequencing, scientists gained the ability to
elucidate genetic information from any given biological system. This technology has become widely
adopted in laboratories around the world, yet has always been hampered by inherent limitations in
throughput, scalability, speed, and resolution that often preclude scientists from obtaining the essential
information they need for their course of study. To overcome these barriers, an entirely new technology
was required—Next-Generation Sequencing (NGS), a fundamentally different approach to sequencing
that triggered numerous ground-breaking discoveries and ignited a revolution in genomic science.
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.
Gene mapping | Genetic map | Physical Map | DNA Data Analysis (upgraded)NARC, Islamabad
Genes are useful markers but not ideal.
Mapped feature that are not genes are called DNA markers.
DNA markers must have at least two alleles to be useful.
DNA sequence features that satisfy this requirement are-
– Restriction Fragment Length Polymorphism (RFLP)
Southern hybridization
PCR
– Simple Sequence Length Polymorphism (SSLP)
– Single Nucleotide Polymorphism (SNP)
Mapping- determining the location of elements with in a genome, with respect to identifiable land marks.
Gene mapping describes the methods used to identify the locus of a gene and the distances between genes.
In simple mapping of genes to specific locations on chromosomes.
Two types
Genetic map
Physical Map
They are useful in predicting results of dihybrid and trihybrid crosses.
It allows geneticists to understand the overall complexity and genetic organization of a particular species.
Identify genes responsible for diseases.
Identify genes responsible for traits.
genetic maps are useful from an evolutionary point of view.
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.
Presented by Dr. Miller at the 40th Annual Symposium "Diagnostic and Clinical Challenges of 20th Century Microbes", held on Nov 18, 2010 in Philadelphia.
diagnosis of cancer, bioluminescent detection, diagnosis of cancer, haplotype mapping, imaging gene expression in vivo, types of cancer diagnosis method, ultrasound imaging
Fluorescence in situ hybridization (FISH) is a cytogenetic technique developed in the early 1980s. FISH uses fluorescent DNA probes to target specific chromosomal locations within the nucleus, resulting in colored signals that can be detected using a fluorescent microscope
this is a presentation on molecular markers that include what is molecular marker, it's types, biochemical markets (alloenzyme), it's classification, data analysis and it's applications
To understand the basic concept of blotting techniques (Southern, northern, western)
To know the main applications and advantages of each of the main types of blotting techniques
To be familiar with the steps (in brief) for performing a blotting procedure
To understand the major similarities & differences between different blotting techniques
To be introduced to an example of applying a blotting technique in diagnosis of diseases (SCA)
DNA Fingerprinting & its techniques by Shiv Kalia (M.Pharma in Analytical Che...Shiv Kalia
DNA fingerprinting and below mention content widely cover in this presentation
History & Introduction of DNA fingerprinting
How was the first DNA fingerprint produced?
Types of DNA Based Markers
Polymerase Chain Reaction (PCR)
PCR based Methodology of DNA fingerprinting
Electrophoresis
Utility of DNA Based Markers
Various DNA Fingerprinting Techniques Advantages & Disadvantages
Authentication of Various Ayurvedic Herbs by DNA Fingerprinting
Advantages of DNA fingerprinting in Plants
Disadvantages of DNA fingerprinting in Plants
CONCLUSION
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.
Comparing Evolved Extractive Text Summary Scores of Bidirectional Encoder Rep...University of Maribor
Slides from:
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
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.
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
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.
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
PRESENTATION ABOUT PRINCIPLE OF COSMATIC EVALUATION
Fluorescence In Situ Hybridization (FISH) Technique in Detection of Plant Pathogens
1. Fluorescence In Situ Hybridization (FISH)
Technique
in Detection of Plant Pathogens
Topic of Doctoral Seminar-I
Presented by
Prashant Waghrulkar,
200136006
Ph.D. Agri. (Plant Pathology),
2nd Year, 3rd Semester
College of Agriculture,
JNKVV, Jabalpur Academic Year: 2021-22
2. Introduction
• It has been estimated that out of 36.5% average total crop losses . . .
• 70-80% of these losses were caused by Fungi
Crop losses (%) Causes
14.1 Diseases
10.2 Insects
12.2 Weeds
• More than 40% of crop loss is due to viral infections
3. Current Methods for Crop Disease Detection
Direct Detection Methods
• Polymerase Chain Reaction (PCR)
• Enzyme-Linked Immunosorbent Assay (ELISA)
• Immunofluorescence (IF)
• Flow Cytometry (FCM)
• Fluorescence in-situ Hybridization (FISH)
Indirect Detection Methods
• Thermography
• Fluorescence Imaging
• Hyperspectral Techniques
• Gas Chromatography
Detection of Plant Diseases Using Portable Sensors
• Biosensor Platforms Based on Nanomaterials
• Affinity Biosensors
- Antibody-Based Biosensors
- DNA/RNA-Based Affinity Biosensor
• Enzymatic Electrochemical Biosensors
• Bacteriophage-Based Biosensors
4. History of ISH
1953:
Watson & Crick demonstrated how the 2 strands of DNA are hold by
hydrogen bonding.
1969:
Gall & Pardue performed the first in situ hybridization experiment.
1991:
First application of FISH to plant cytogenetics by Leitch et al.
Joseph Gall Mary-Lou Pardue
5. Fluorescence In Situ Hybridization (FISH)
• It is a molecular detection technique which include hybridization of
DNA probes & target gene followed by microscopy.
• In this technique fluorescent probes bind to particular parts of
a nucleic acid sequence with a high degree of
sequence complementarity.
6. Probe: Single strand of DNA or RNA that is complementary to a
nucleotide sequence of interest.
• It is approx. 20-50 oligonucleotide pairs & covering a space of 40–50
bp.
• A wide range of probes, extending from whole genomes to small
cloned probes (1–10 kb), can be used.
There are basically 3 types of probes
• Whole chromosome painting probes
• Repetitive sequence probes
• locus specific probes
Probe Labelling (Tagging by Nick translation)
• DNase1 nick the one strand of DNA & break the phosphodieaster
bond then some of the nucleotides of a DNA sequence replaced with
label.
• It can also be used for Radiolabeling.
8. Applications
• To detect & localize the presence or absence of
specific DNA sequences on chromosomes.
• To detect chromosomal aberrations (additions, deletions, insertions,
inversions, copy number, translocations).
• Used for gene mapping.
• Karyotyping by Chromosome painting
Types of chromosome painting
Whole chromosome
painting probe
(WPP)
Chromosome terminal
band painting probes
(TPP)
Chromosome arm
painting probes
(APP)
Chromosome
enumeration probes
(CEP)
Cont...
10. • To detect & localize specific RNA targets (mRNA: messenger
RNA, lncRNA: Long non-coding RNA & miRNA: micro RNA) in cells,
circulating tumor cells, & tissue samples.
• It help to define the spatial-temporal patterns of gene
expression within cells and tissues.
Central DOGMA
DNA to RNA to Protein
rRNA
Large subunit
Small subunit
tRNA
mRNA
RNA
Ribosomal RNA
Messenger RNA
Transfer RNA
Amino acid delivered by tRNA form a
Chain, then fold & form a protein
Applications
Cont...
11. • Used in prenatal diagnosis of diseases like Downy syndrome, cancer
etc.
• Helps in genetic counseling, medicine, & species identification.
• Due to the presence of pathogen-specific ribosomal RNA (rRNA)
sequences in plants, recognizing this specific information by FISH can
help detect the pathogen infections.
• In addition to bacterial pathogens, FISH could also be used to detect
fungi & viruses and other endosymbiotic bacteria that infect the
plant.
Applications
13. Transmitted light Actin in Green
Actin is a microfilamentous protein of cytoskeleton
& important for shape of the cell
Mitochondria in Orange Nucleus in Blue
14. Advantages
• High Affinity & Specificity of DNA probes provide high single-cell sensitivity
because the probe will bind to each of the ribosomes in the sample.
• Can be detect culturable & yet-to-be cultured (so called unculturable)
organisms in order to investigate complex microbial communities.
Limitations
Detection Limit
Lies in the range of around 103 CFU/mL.
Accuracy & reliability
Highly dependent on specificity of nucleotide probes. Probe designing is
challenging.
Cause of false negative results
Insufficient penetration, higher order structure of target or probe (e.g., 3D
rRNA, loop & hairpin formation), low rRNA content, & photobleaching.
16. Limitations
Autofluorescence (false positive results)
Natural fluorescence
• Lipofuscins: A native autofluorescent material in certain large neurons in the
CNS.
• Elastin: It found in blood vessel walls & contain a similar fluorophore that
found in collagen.
• Collagen: Extracellular material between the smooth muscle and in the
adventitia layer.
Fixative-induced Fluorescence: Aldehyde, Glutaraldehyde
Cont...
Fang Y, Ramasamy RP. Current and Prospective Methods for Plant Disease Detection. Biosensors (Basel). 2015;5(3):537-561. Published 2015 Aug 6. doi:10.3390/bios5030537
Fang Y, Ramasamy RP. Current and Prospective Methods for Plant Disease Detection. Biosensors (Basel). 2015;5(3):537-561. Published 2015 Aug 6. doi:10.3390/bios5030537
Autofluorescence: Causes and Cures. Toronto Western Research Institute, University Health Work
Autofluorescence: Causes and Cures. Toronto Western Research Institute, University Health Work
zeiss-campus.magnet.fsu.edu/articles/spectralimaging/introduction.html
