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.
Topics included - Introduction; explanation; examples like blue white screening method, antibiotic resistance; Extra information regarding - detection of oncogene in vertebrates and role of sleeping beauty; Merits and demerits of insertional inactivation.
Topics included - Introduction; explanation; examples like blue white screening method, antibiotic resistance; Extra information regarding - detection of oncogene in vertebrates and role of sleeping beauty; Merits and demerits of insertional inactivation.
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.
Blotting technique including Southern , Northern and Western blotting Rohit Mondal
he given ppt contains all the blotting techniques which is being studied by students in Biotechnology related subject and this PPT contais all blotting techniques in a very elaborative concise manner includes procedure principle application etc so which itwould help any bio student to take proper knowledge in this topic. I hope you will enjoy the content of the topic and would be able to grasp the topic properly
Making of preparations involves different techniques, shall train a learner to be competant enough to know the importance of solution in different dosage form.
It will make learners understand the borderline concepts related with different system, shall train a students to be competent enough to know about the same.
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
Follow us on: Pinterest
Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
Lung Cancer: Artificial Intelligence, Synergetics, Complex System Analysis, S...Oleg Kshivets
RESULTS: Overall life span (LS) was 2252.1±1742.5 days and cumulative 5-year survival (5YS) reached 73.2%, 10 years – 64.8%, 20 years – 42.5%. 513 LCP lived more than 5 years (LS=3124.6±1525.6 days), 148 LCP – more than 10 years (LS=5054.4±1504.1 days).199 LCP died because of LC (LS=562.7±374.5 days). 5YS of LCP after bi/lobectomies was significantly superior in comparison with LCP after pneumonectomies (78.1% vs.63.7%, P=0.00001 by log-rank test). AT significantly improved 5YS (66.3% vs. 34.8%) (P=0.00000 by log-rank test) only for LCP with N1-2. Cox modeling displayed that 5YS of LCP significantly depended on: phase transition (PT) early-invasive LC in terms of synergetics, PT N0—N12, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), G1-3, histology, glucose, AT, blood cell circuit, prothrombin index, heparin tolerance, recalcification time (P=0.000-0.038). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and PT early-invasive LC (rank=1), PT N0—N12 (rank=2), thrombocytes/CC (3), erythrocytes/CC (4), eosinophils/CC (5), healthy cells/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), stick neutrophils/CC (9), monocytes/CC (10); leucocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
CONCLUSIONS: 5YS of LCP after radical procedures significantly depended on: 1) PT early-invasive cancer; 2) PT N0--N12; 3) cell ratio factors; 4) blood cell circuit; 5) biochemical factors; 6) hemostasis system; 7) AT; 8) LC characteristics; 9) LC cell dynamics; 10) surgery type: lobectomy/pneumonectomy; 11) anthropometric data. Optimal diagnosis and treatment strategies for LC are: 1) screening and early detection of LC; 2) availability of experienced thoracic surgeons because of complexity of radical procedures; 3) aggressive en block surgery and adequate lymph node dissection for completeness; 4) precise prediction; 5) adjuvant chemoimmunoradiotherapy for LCP with unfavorable prognosis.
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
NYSORA Guideline
2 Case Reports of Gastric Ultrasound
Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Ve...kevinkariuki227
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
The prostate is an exocrine gland of the male mammalian reproductive system
It is a walnut-sized gland that forms part of the male reproductive system and is located in front of the rectum and just below the urinary bladder
Function is to store and secrete a clear, slightly alkaline fluid that constitutes 10-30% of the volume of the seminal fluid that along with the spermatozoa, constitutes semen
A healthy human prostate measures (4cm-vertical, by 3cm-horizontal, 2cm ant-post ).
It surrounds the urethra just below the urinary bladder. It has anterior, median, posterior and two lateral lobes
It’s work is regulated by androgens which are responsible for male sex characteristics
Generalised disease of the prostate due to hormonal derangement which leads to non malignant enlargement of the gland (increase in the number of epithelial cells and stromal tissue)to cause compression of the urethra leading to symptoms (LUTS
micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
2. Objective:
To make students understand regarding the Genetic
Engineering: Recombinant DNA technology with
respect to its methods for gene transfer, & different
Genetic Engineering techniques.
Outcome:
Students are familiar with the different DNA
transfer techniques & these method involves
separation, transfer and hybridization
3. What is blotting?
Blots are techniques for transferring DNA ,
RNA and proteins onto a carrier so they can
be separated, and often follows the use of a
gel electrophoresis. The Southern blot is used
for transferring DNA, the Northern blot for
RNA and the western blot for PROTEIN.
5. SOUTHERN BLOTTING
Professor Sir Edwin Southern,
Professor of Biochemistry and Fellow
of Trinity developed this method in
1975.
Southern won the Lasker Award for
Clinical Medical Research prize for
the method of finding specific DNA
sequences he developed this procedure
at Edinburgh University more than 30
years ago. The technique is known as Professor Sir Edwin Southern
6. Cont….
This method Involves separation, transfer and
hybridization.
It is a method routinely used in molecular biology
for detection of a specific DNA sequence in DNA
samples. The DNA detected can be a single gene, or
it can be part of a larger piece of DNA such as a viral
genome.
7. Cont….
Southern blotting combines agarose gel electrophoresis
for size separation of DNA with methods to transfer the
size separated DNA to a filter membrane for probe
hybridization.
The key to this method is Hybridization.
Hybridization - Process of forming a double-stranded
DNA molecule between a single-stranded DNA probe and
a single-stranded target patient DNA.
8. PRINCIPLE
1. The mixture of molecules is separated.
2. The molecules are immobilized on a matrix.
3. The probe is added to the matrix to bind to the
molecules.
4. Any unbound probes are then removed.
5. The place where the probe is connected corresponds
to the location of the immobilized target molecule.
12. Steps in Southern Blotting
1. Digest the DNA with an
appropriate restriction
enzyme.
2.The complex mixture of
fragments is subjected to gel
electrophoresis to separate
the fragments according to
size.
13. Cont….
3.The restriction fragments
present in the gel are
denatured with alkali and
transferred onto
4. a nitrocellulose filter or
nylon membrane by
blotting.
This procedure preserves
the distribution of the
fragments in the gel,
14. Cont….
5.The filter is incubated under
hybridization conditions with a
specific radiolabeled DNA
probe.
The probe hybridizes to the
complementary DNA
restriction fragment.
15. Cont….
6. Excess probe is washed away and
the probe bound to the filter is
detected by autoradiography,
which reveals the DNA fragment
to which the probe hybridized.
16.
17.
18. APPLICATIONS
Southern blots are used in gene discovery ,
mapping, evolution and development studies,
diagnostics and forensics (It is used for DNA
fingerprinting, preparation of RFLP maps)
Identification of the transferred genes in
transgenic individuals, etc.
19. Southern blots allow investigators to determine the
molecular weight of a restriction fragment and to
measure relative amounts in different samples.
Southern blot is used to detect the presence of a
particular bit of DNA in a sample
Analyze the genetic patterns which appear in a
person's DNA.
20. Northern Blotting
Northern blotting is a technique for detection of
specific RNA sequences. Northern blotting was
developed by James Alwine and George Stark at
Stanford University (1979) and was named such by
analogy to Southern blotting
21. Steps involved in Northern blotting
1. RNA is isolated from several
biological samples (e.g.
various tissues, various
developmental stages of
same tissue etc.)
* RNA is more susceptible
to degradation than DNA.
22. Cont……
2. Sample’s are loaded on
gel and the RNA samples
are separated according to
their size on an agarose
gel .
The resulting gel following
after the electrophoresis
23. Cont……
3. The gel is then blotted
on a nylon membrane
or a nitrocellulose
filter paper by creating
the sandwich
arrangement.
24. Cont……
4. The membrane is placed in a
dish containing hybridization
buffer with a labeled probe.
Thus, it will hybridize to the
RNA on the blot that
corresponds to the sequence of
interest.
5. The membrane is washed to
25. Cont……
6. The labeled probe is detected via
autoradiography or via a
chemiluminescence reaction (if a
chemically labeled probe is
used). In both cases this results in
the formation of a dark band on an
X-ray film.
Now the expression patterns of the
sequence of interest in the different
samples can be compared.
26. APPLICATIONS
A standard for the study of gene expression at the
level of mRNA (messenger RNA transcripts)
Detection of mRNA transcript size
Study RNA degradation
Study RNA splicing
Study RNA half-life
Often used to confirm and check transgenic /
knockout mice (animals)
27. Disadvantage of Nourthern plotting
1.The standard northern blot method is relatively less
sensitive than nuclease protection assays and RT-
PCR
2. Detection with multiple probes is a problem
3. If RNA samples are even slightly degraded by RNases,
the quality of the data and quantitation of expression
is quite negatively affected.
28. Western blotting
Western blotting (1981) is an Immunoblotting
technique which rely on the specificity of binding
between a protein of interest and a probe (antibody
raised against that particular protein) to allow
detection of the protein of interest in a mixture of
many other similar molecules.
The SDS PAGE technique is a prerequisite for
29. Steps in western blotting
1. A protein sample is
subjected to
electrophoresis on an SDS-
polyacrylamide gel.
2. Electroblotting transfers
the separated proteins from
the gel to the surface of a
30. Cont…
3. The blot is incubated with a generic protein
(such as milk proteins or BSA) which binds
to any remaining sticky places on the
nitrocellulose.
4. An antibody that is specific for the protein of
interest (the primary antibody - Ab1) is
added to the nitrocellulose sheet and reacts
with the antigen. Only the band containing
the protein of interest binds the antibody,
31. Cont…
5. After washing for removal of non-
specifically bound Ab1, second
antibody (Ab2)is added, which
specifically recognizes the Fc
domain of the primary antibody and
binds it. Ab2 is radioactively
labeled, or is covalently linked to a
reporter enzyme, which allows to
visualize the protein-Ab1-Ab2
33. Applications
1. The confirmatory HIV test
2. Western blot is also used as the definitive test for
Bovine spongiform encephalopathy (BSE(
3. Some forms of Lyme disease testing employ
Western blotting .
34. What Do you Think about following………..
Q.1.What do you mean by Blotting Techniques?
Q.2. Give detail explanation about Southern, Western
&
Northern Blotting Techniques?
Q.3. Give applications of each techniques?