published a DNA sequencing method in 1977 based on chemical modification of DNA and subsequent cleavage at specific bases. Also known as chemical sequencing, this method allowed purified samples of double-stranded DNA to be used without further cloning.
Maxam-Gilbert sequencing requires radioactive labeling at one 5' end of the DNA and purification of the DNA fragment to be sequenced. Chemical treatment then generates breaks at a small proportion of one or two of the four nucleotide bases in each of four reactions (G, A+G, C, C+T). The concentration of the modifying chemicals is controlled to introduce on average one modification per DNA molecule. Thus a series of labeled fragments is generated, from the radiolabeled end to the first "cut" site in each molecule. The fragments in the four reactions are electrophoresed side by side in denaturing acrylamide gels for size separation. To visualize the fragments, the gel is exposed to X-ray film for autoradiography, yielding a series of dark bands each corresponding to a radiolabeled DNA fragment, from which the sequence may be inferred.
Open reading frame is part of reading frame that contains no stop codons or region of amino acids coding triple codons.
ORF starts with start codon and ends at stop codon.
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.
This is technique used widely for protein separation from a mixture and is very easy and less costly method. Slides cover all essential points about EMSA and it is quite interesting to know that how it detect and separate different proteins and their mobility shift assay.
Open reading frame is part of reading frame that contains no stop codons or region of amino acids coding triple codons.
ORF starts with start codon and ends at stop codon.
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.
This is technique used widely for protein separation from a mixture and is very easy and less costly method. Slides cover all essential points about EMSA and it is quite interesting to know that how it detect and separate different proteins and their mobility shift assay.
Sanger sequencing is one of the DNA sequencing methods used to identify and determine the sequence (Nucleotide) of DNA .This is an enzymatic method of sequencing developed by Fred Sanger.
Scoring system is a set of values for qualifying the set of one residue being substituted by another in an alignment.
It is also known as substitution matrix.
Scoring matrix of nucleotide is relatively simple.
A positive value or a high score is given for a match & negative value or a low score is given for a mismatch.
Scoring matrices for amino acids are more complicated because scoring has to reflect the physicochemical properties of amino acid residues.
Genomic library and shotgun sequencing. It includes the topics about genomic library,construction method, its uses and applications, shotgun sequencing, difference between random and whole genome sequencing, its advantages and disadvantages etc.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
DNA Sequencing : Maxam Gilbert and Sanger SequencingVeerendra Nagoria
DNA sequencing is a technique to find out the exact arrangement of Nucleotides to make one strand of DNA. DNA sequencing helps in numerous ways from sequence information to paternity testing, mutation detection etc. Traditionally two approaches were used to solve the problem. First is based of enzymes and Second is based on ddNTPs to sequence the DNA using gel electrophoresis technique.
Sanger sequencing is a method of DNA sequencing based on the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication.
Sanger sequencing is one of the DNA sequencing methods used to identify and determine the sequence (Nucleotide) of DNA .This is an enzymatic method of sequencing developed by Fred Sanger.
Scoring system is a set of values for qualifying the set of one residue being substituted by another in an alignment.
It is also known as substitution matrix.
Scoring matrix of nucleotide is relatively simple.
A positive value or a high score is given for a match & negative value or a low score is given for a mismatch.
Scoring matrices for amino acids are more complicated because scoring has to reflect the physicochemical properties of amino acid residues.
Genomic library and shotgun sequencing. It includes the topics about genomic library,construction method, its uses and applications, shotgun sequencing, difference between random and whole genome sequencing, its advantages and disadvantages etc.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
DNA Sequencing : Maxam Gilbert and Sanger SequencingVeerendra Nagoria
DNA sequencing is a technique to find out the exact arrangement of Nucleotides to make one strand of DNA. DNA sequencing helps in numerous ways from sequence information to paternity testing, mutation detection etc. Traditionally two approaches were used to solve the problem. First is based of enzymes and Second is based on ddNTPs to sequence the DNA using gel electrophoresis technique.
Sanger sequencing is a method of DNA sequencing based on the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication.
DNA sequencing is the process of determining the sequence of nucleotides (A, T, G, and C) in the DNA. It includes method or technology that is used to determine the order of the four bases: adenine, thymine, guanine and cytosine.
Los días 20 y 21 de octubre de 2016, la Fundacion Ramón Areces organizó un simposio internacional para analizar las 'Enfermedades raras de la piel: de la clínica al gen y viceversa'. El doctor Fernando Larcher Laguzzi, del CIEMAT-Universidad Carlos III de Madrid-IIS Fundación Jiménez Díaz, ejerció de coordinador.
A detailed description about the basic steps involved in the - PCR - Polymerase Chain Reaction, its applications,its limitations and steps to overcome it.
The chain-termination method developed by Frederick Sanger and coworkers in 1977. This method used fewer toxic chemicals and lower amounts of radioactivity than the Maxam and Gilbert method. Because of its comparative ease, the Sanger method was soon automated and was the method used in the first generation of DNA sequencers.
A chart showing the fate of each part of an early embryo, in a particular blastula stage is called fate maps. It is done because the correct interpretation of gastrulation is impossible without the knowledge of the position which are the presumptive germinal layers (Ectoderm, Mesoderm and Endoderm) occupy in blastula.
Fate mapping is a method used in developmental biology to study the embryonic origin of various adult tissues and structures. The "fate" of each cell or group of cells is mapped onto the embryo, showing which parts of the embryo will develop into which tissue. When carried out at single-cell resolution, this process is called cell lineage tracing. It is also used to trace the development of tumors.
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).
Bacteriophage- types, structure and morphology of t4 phage, morphogenesisDr. Dinesh C. Sharma
Escherichia virus T4 is a species of bacteriophages that infect Escherichia coli bacteria. It is a member of virus subfamily Tevenvirinae (not to be confused with T-even bacteriophages, which is an alternate name of the species). T4 is capable of undergoing only a lytic lifecycle and not the lysogenic lifecycle.
Each cell of a multicellular organism contain the same genetic material, but the expression of the gene is different in different type of cell group. On the basis of expression requirement they are grouped in to
Structural Gene- Mostly expressed once in a life
Vital Gene- Involved in of vital biochemical processes such as respiration and need to be expressed all the time
Functional Gene- Genes are not expressed all the time. They are switched on an off at need
The regulation of Gene required in case of functional gene and its explained by Francois Jacob, Jacques Monod and Andre Lwoff (Nobal Prize in 1961)
From studies and predictions such as Dreyer and Bennett's, it shows that the light chains and heavy chains are encoded by separate multigene families on different chromosomes. They are referred to as gene segments and are separated by non-coding regions. The rearrangement and organization of these gene segments during the maturation of B cells produce functional proteins. The entire process of rearrangement and organization of these gene segments is the vital source where our body immune system gets its capabilities to recognize and respond to variety of antigens.
The cells of the B line synthesize immunoglobulins. They are either produced at a membrane (on the surface of the B-lymphocytes) or are secreted (by the plasmocytes)
Theory of preformation,
Epigenetic theory,
Theory of pengenesis,
Recapitulation theory,
Germplasm theory,
Mosaic theory,
Regulated theory,
Gradient theory
Theory of organizers.
Sericulture is the cultivation of silkworms to produce silk. Bombyx mori (the caterpillar of the domesticated silk moth) is the most widely used species of silkworms.
A device that computes, especially a programmable electronic machine that performs high-speed mathematical or logical operations or that assembles, stores, correlates, or otherwise processes information.
As a result of our consumer culture lifestyle, we are polluting the earth and slowly changing its temperature. As a result, weather patterns will be less predictable and water level will rise significantly
Climate change is an extended change in the Earth’s regular pattern of atmospheric conditions and its fluctuations
Global warming is caused by an enhanced greenhouse effect mostly caused by anthropogenic activity
- 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
Factory Supply Best Quality Pmk Oil CAS 28578–16–7 PMK Powder in Stockrebeccabio
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MANAGEMENT OF ATRIOVENTRICULAR CONDUCTION BLOCK.pdfJim Jacob Roy
Cardiac conduction defects can occur due to various causes.
Atrioventricular conduction blocks ( AV blocks ) are classified into 3 types.
This document describes the acute management of AV block.
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.
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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
Anti ulcer drugs and their Advance pharmacology ||
Anti-ulcer drugs are medications used to prevent and treat ulcers in the stomach and upper part of the small intestine (duodenal ulcers). These ulcers are often caused by an imbalance between stomach acid and the mucosal lining, which protects the stomach lining.
||Scope: Overview of various classes of anti-ulcer drugs, their mechanisms of action, indications, side effects, and clinical considerations.
ARTIFICIAL INTELLIGENCE IN HEALTHCARE.pdfAnujkumaranit
Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
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
Title: Sense of Smell
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 primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
1. By-
Dr. Dinesh C. Sharma
Head, Zoology
K.M. Govt. Girls P. G. College
Badalpur, G.B. Nagar
dr_dineshsharma@hotmail.com
“the process of determining the sequence of nucleotides (A, T, G, and C) in a piece of DNA”
3. Maxam-Gilbert sequencing published a DNA sequencing
method in 1977 based on chemical modification of DNA and
subsequent cleavage at specific bases. Also known as chemical
sequencing, this method allowed purified samples of double-
stranded DNA to be used without further cloning.
Maxam-Gilbert sequencing requires radioactive labeling at one 5' end
of the DNA and purification of the DNA fragment to be sequenced.
Chemical treatment then generates breaks at a small proportion of one
or two of the four nucleotide bases in each of four reactions (G, A+G,
C, C+T). The concentration of the modifying chemicals is controlled
to introduce on average one modification per DNA molecule. Thus a
series of labeled fragments is generated, from the radiolabeled end to
the first "cut" site in each molecule. The fragments in the four reactions
are electrophoresed side by side in denaturing acrylamide gels for size
separation. To visualize the fragments, the gel is exposed to X-ray film
for autoradiography, yielding a series of dark bands each corresponding
to a radiolabeled DNA fragment, from which the sequence may be
inferred.
4. Maxam–Gilbert sequencing requires radioactive labeling at
one 5′ end of the DNA fragment to be sequenced (typically by
a kinase reaction using gamma-32P ATP) and purification of
the DNA.
Chemical treatment generates breaks at a small proportion of
one or two of the four nucleotide bases in each of four
reactions (G, A+G, C, C+T). For example, the purines (A+G)
are depurinated using formic acid, the guanines (and to some
extent the adenines) are methylated by dimethyl sulfate, and
the pyrimidines (C+T) are hydrolysed using hydrazine. The
addition of salt (sodium chloride) to the hydrazine reaction
inhibits the reaction of thymine for the C-only reaction. The
modified DNAs may then be cleaved by hot piperidine;
(CH2)5NH at the position of the modified base.
The concentration of the modifying chemicals is controlled to introduce on average
one modification per DNA molecule. Thus a series of labeled fragments is
generated, from the radiolabeled end to the first "cut" site in each molecule.
The fragments in the four reactions are electrophoresed side by side in denaturing
acrylamide gels for size separation. To visualize the fragments, the gel is exposed to
X-ray film for autoradiography, yielding a series of dark bands each showing the
location of identical radiolabeled DNA molecules. From presence and absence of
certain fragments the sequence may be inferred
6. 5”- C T C G A G T G T A T C G
A C -3”
3”- T C T C T C A C A T A G C
T G -5”
| | | | | | | | | | | | | | |
1-Denturation @ 95O C
5”- C T C G A G T G T A T C G
A C -3”
3”- T C T C T C A C A T A G C
T G -5”
Four samples (A+G), G, (T+C) and C
A+G G T+C C
8. 1
A+G
2
G
3
T+C
4
C
B-Add Formic Acid in 1: Formic acid breaks link
between a purine (A+G) and the deoxyribose to which it attached
5”- C T C G A G T G T A T C G
A C -3”
5”- C T
C
5”- C T C G A
G T
5”- C T C G
A
5”- C T C
G
5”- C T C G A G T
G T5”- C T C G A G T G T
A T C5”- C T C G A G T G T A T
7
Fragmen
t
9. 1
A+G
2
G
3
T+C
4
C
C-Dimethyal Sulfate in 2: Methylation of
Guanine by DMS
5”- C T C G A G T G T A T C G
A C -3”
5”- C T C
5”- C T C G A G T G T A
T C
5”- C T C G A G T
5”- C T C G A
4
Fragmen
t
10. 1
A+G
2
G
3
T+C
4
C
D-Add Hydrazine in 3: The pyrimidines are
hydrolyzed by using hydrazine
5”- C T C G A G T G T A T C G
A C -3”5”- C T C G A G T G T A T C G
A C5”- C T C G A G T G T A T C G
A5”- C T C G A G T G T A T
5”- C T C G A G T G T A
5”- C T C G A G T G
5”- C T C G A G
5”- C T
5”- C
8
Fragmen
t
11. 1
A+G
2
G
3
T+C
4
C
E-Add Hydrazine and NaCl in 4: The addition of
NaCl to hydrazine reaction inhibits the reaction of thymine for –C
only reaction
5”- C T C G A G T G T A T C G
A C -3”
5”- C
5”- C T C G A G T G T A T C G
A
5”- C T C G A G T G T A T
5”- C T
4
Fragmen
t
12. 1
A+G
2
G
3
T+C
4
C
F-Add Piperidine in all 4: The modified DNAs
cleaved by hot piperidine at the position of the modified base
5”- C T C G A G T G T A T C G
A C -3”
14. • The negative charge of phosphate
backbone move the DNA
fragments towards the positively
charged anode
• Smaller DNA fragments migrate
more rapidly than larger DNA
fragments
Small
Large
18. 5”- C T
C
5”- C T C G A G
T
5”- C T C G A
5”- C T C G
5”- C T C G A G T G
T
5”- C T C G A G T G T A T C
5”- C T C G A G T G T A T C G
7
19. 5”- C T C
5”- C T C G A G T G T A
T C
5”- C T C G A G T
5”- C T C G A
4
20. 5”- C T C G A G T G T A T C
G A C
5”- C T C G A G T G T A T C
G A
5”- C T C G A G T G T A T
5”- C T C G A G T G T A
5”- C T C G A G T G
5”- C T C G A G
5”- C T
5”- C
8
21. 5”- C
5”- C T C G A G T G T A T
C G A
5”- C T C G A G T G T A T
5”- C T
4
22. 5”- C T C
5”- C T C G A G T
5”- C T C G A
5”- C T C G
5”- C T C G A G T
G T
5”- C T C G A G T G T A T
C
5”- C T C G A G T G T A T
C G
5”- C T C
5”- C T C G A G T G T A
T C
5”- C T C G A G T
5”- C T C G A
5”- C T C G A G T G T A T C
G A C
5”- C T C G A G T G T A T C
G A
5”- C T C G A G T G T A T
5”- C T C G A G T G T A
5”- C T C G A G T G
5”- C T C G A G
5”- C T
5”- C 5”- C
5”- C T C G A G T G T A T C
G A
5”- C T C G A G T G T A T
5”- C T
C
C
C
C
T
T
T
T
G
G
G
G
A
A
A
C
A
G
C
T
A
T
G
T
G
A
G
C
T
C