Last summer I interned at the Genetics Laboratory, Department of Assisted Reproduction and Genetics, Jaslok Hospital. Here is a powerpoint of my hands on experience of karyotyping blood sample from the stage of culture set-up to chromosome analysis
Define karyotype and FISH
Describe the procedure of karyotyping and FISH
Explain chromosomal abnormalities through karyotyping and FISH
Describe the principles of FISH
Presented by-
Dr. Subarna Das
Resident, MS Anatomy
Phase-A, Year-1, Block-2
Guided by-
Prof. Laila Anjuman Banu
Chairman
Department of Anatomy, BSMMU
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.
Define karyotype and FISH
Describe the procedure of karyotyping and FISH
Explain chromosomal abnormalities through karyotyping and FISH
Describe the principles of FISH
Presented by-
Dr. Subarna Das
Resident, MS Anatomy
Phase-A, Year-1, Block-2
Guided by-
Prof. Laila Anjuman Banu
Chairman
Department of Anatomy, BSMMU
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.
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.
Chromosomes count plays an important role cytogenetics and breeding. As a breeder of citrus, this is one of MSc degree experiment at University of Miyazaki, Japan.
my lab presentation
Southern, Northern and Western Blotting methods in genetic EngineeringRavi Raj
Blotting is a method in which a macromolecule is immobilized on a blotting matrix and subsequently probed with a detectable ligand to determine whether the macromolecule binds that specific ligand. The immobilized macromolecule can be DNA, RNA or protein, in which case one generates DNA blots (Southern blots), RNA blots (Northern blots) (1), or protein blots (Western blots).
Flow Cytometry Training talks - part 1
This forms the first session of the Garvan Flow , Flow Cytometry Training course. this is a 1 1/2 day training course aimed at giving new and experienced researchers a better understanding of cytometry in medical and biological research.
Techniques of Assessment of Genetic Changes Saranya Roy
the various techniques used for the assessment of genetic changes in humans, including PCR, Northern and Southern blotting techniques and their applications
Development of an Automated Comet Assay for Genotoxicity Assessment on TK6 ce...HCS Pharma
Quantifying DNA damage is mandatory to assess potential adverse effects of candidate drugs or molecules or extracts developed in the dermo-cosmetic industry, but also to assess the efficacy of therapeutic approaches with the aim of producing tumor cell genotoxicity in cancer treatment.
The comet assay is a sensitive, well established technique for quantifying DNA damage in eukaryotic cells. Compatible with the detection of a wide range of DNA damaging agents, its principle consists in the migration of fragmented DNA in an electrophoresis gel (damaged DNA forming the tail of the comet), while intact DNA moves at a slower rate (head of the comet). The percentage of fragmented DNA in the comet tail is a direct measure of DNA damage.
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.
Chromosomes count plays an important role cytogenetics and breeding. As a breeder of citrus, this is one of MSc degree experiment at University of Miyazaki, Japan.
my lab presentation
Southern, Northern and Western Blotting methods in genetic EngineeringRavi Raj
Blotting is a method in which a macromolecule is immobilized on a blotting matrix and subsequently probed with a detectable ligand to determine whether the macromolecule binds that specific ligand. The immobilized macromolecule can be DNA, RNA or protein, in which case one generates DNA blots (Southern blots), RNA blots (Northern blots) (1), or protein blots (Western blots).
Flow Cytometry Training talks - part 1
This forms the first session of the Garvan Flow , Flow Cytometry Training course. this is a 1 1/2 day training course aimed at giving new and experienced researchers a better understanding of cytometry in medical and biological research.
Techniques of Assessment of Genetic Changes Saranya Roy
the various techniques used for the assessment of genetic changes in humans, including PCR, Northern and Southern blotting techniques and their applications
Development of an Automated Comet Assay for Genotoxicity Assessment on TK6 ce...HCS Pharma
Quantifying DNA damage is mandatory to assess potential adverse effects of candidate drugs or molecules or extracts developed in the dermo-cosmetic industry, but also to assess the efficacy of therapeutic approaches with the aim of producing tumor cell genotoxicity in cancer treatment.
The comet assay is a sensitive, well established technique for quantifying DNA damage in eukaryotic cells. Compatible with the detection of a wide range of DNA damaging agents, its principle consists in the migration of fragmented DNA in an electrophoresis gel (damaged DNA forming the tail of the comet), while intact DNA moves at a slower rate (head of the comet). The percentage of fragmented DNA in the comet tail is a direct measure of DNA damage.
Cloning is the process of producing genetically identical individuals of an organism either naturally or artificially.
It is the process of taking genetic information from one living thing and creating identical copies of it. The copied material is called a clone.
Nature has been doing it for millions of years. For example, identical twins have almost identical DNA, and asexual reproduction in some plants and organisms can produce genetically identical offspring.
Cloning in biotechnology refers to the process of creating clones of organisms or copies of cells or DNA fragments (molecular cloning).
This slide gives you details about
1. embalming
2. museum techniques
3. principles of karyotyping
chemicals used for embalming
instruments used for embalming
embalming procedures
uses of embalming
procedures for museum techniques
procedure for storing specimens
instruments used in specimen storage
different types of jars
karyotyping definition
procedure for karyotyping
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
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 .
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
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.
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.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
2. About the Observership
• Location: Genetics Laboratory, Department of Assisted Reproduction and Genetics at
Jaslok Hospital
Tasks Completed
• Observed routine Cytogenetic diagnostic techniques on various human tissues such as
peripheral blood, bone marrow, amniotic fluid and chronic villi from products of conception.
• Hands on experience of karyotyping blood sample from the stage of culture set-up to
chromosome analysis
• Observed FISH (Fluorescence in situ hybridisation) set up, hybridisation, post hybridisation
washes and analysis.
3. Techniques
• Cytogenetics- Study of chromosomes and their abnormalities
• Karyotyping and FISH- used to diagnose genetic diseases.
5. Method for Karyotyping
• Planting: Culture the tissue for appropriate amount of time (usually 48 to 72
hours).
• Harvesting: Add a spindle poison (we used colchicine) to “arrest” the cells in
metaphase. The is because chromosomes are maximally condensed in
metaphase and thus easiest to see. Then add a hypotonic solution to swell up the
cells.
• Washing: Wash the cells
• Dropping: Drop the sample onto the slide from a height
• Banding: Stain with designated nuclear stain. This helps to identify individual
Chromosomes. (Giemsa banding/G banding done in this case)
• Analysing: Capture images of slides under microscope and using software
arrange the chromosomes into a karyotype.
13. FISH
• Fluorescence In Situ Hybridisation
• Take DNA probe that is complementary to the
selected region on the chromosome.
Denature the probe and chromosome
sample.
• Hybridization takes place for 3 hours, or
overnight. Washing is then done. Observe
coloured signals under fluorescent
microscope. Capture image. Chromosome
deletions, aneuploidies (duplications) and
translocations (rearrangements) can be
detected.
• A specific chromosome is looked at, which
gives the observer some idea of which
disease they’re testing for.
14. DNA extraction
• Some diseases can only be
diagnosed on a molecular level
(not via chromosomes). Thus
using the patient’s DNA, one can
test for microdeletions.
• PCR- polymer chain reaction. A
specific gene is located, and that
gene is replicated.
