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.
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.
Principle: The method is characterized by transferring the protein which run on a gel by electrophoresis on to a nitro cellulose membrane.
It is widely used analytical technique in molecular biology to detect specific proteins in a sample of tissue homogenate or extracts.
PAGE is a subtype of the gel electrophoresis whereby the normal gel is replaced with polyacrylamide gels use as the support matrix.
widely used and has very much importance.
COMPLETE PROCEDURE & USES are described in the slide.
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
The technique of molecular biology like DNA isolation, RNA isolation, PCR, Western blot, RFLP, etc was developed with development in science. This presentation includes the method of DNA and RNA isolation and their Quantification techniques.
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.
Principle: The method is characterized by transferring the protein which run on a gel by electrophoresis on to a nitro cellulose membrane.
It is widely used analytical technique in molecular biology to detect specific proteins in a sample of tissue homogenate or extracts.
PAGE is a subtype of the gel electrophoresis whereby the normal gel is replaced with polyacrylamide gels use as the support matrix.
widely used and has very much importance.
COMPLETE PROCEDURE & USES are described in the slide.
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
The technique of molecular biology like DNA isolation, RNA isolation, PCR, Western blot, RFLP, etc was developed with development in science. This presentation includes the method of DNA and RNA isolation and their Quantification techniques.
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.
GPCRs are the most dynamic and most abundant all the receptors. The G protein-coupled receptor (GPCR) superfamily comprises the largest and most diverse group of proteins in mammals. GPCRs are responsible for every aspect of human biology from vision, taste, sense of smell, sympathetic and parasympathetic nervous functions, metabolism, and immune regulation to reproduction. GPCRs interact with a number of ligands ranging from photons, ions, amino acids, odorants, pheromones, eicosanoids, neurotransmitters, peptides, proteins, and hormones.
Nevertheless, for the majority of GPCRs, the identity of their natural ligands is still unknown, hence remain orphan receptors.
The simple dogma that underpins much of our current understanding of GPCRs, namely,
one GPCR gene− one GPCR protein− one functional GPCR− one G protein −one response
is showing distinct signs of wear.
This PPT discusses about the main types of Nucleic Acid Based Techniques - Blotting (Southern, Northen, Western)
Do Leave a comment if you liked the presentation, so that i can improve more and share more!
Southern blotting is a laboratory technique used to detect specific DNA sequences in DNA samples. It involves several steps:
Restriction Digestion: DNA from a biological sample (such as blood or tissue) is broken into smaller fragments using restriction enzymes, which cut the DNA at specific sequences.
Electrophoresis: The DNA fragments are separated based on their molecular weights using gel electrophoresis. This process allows smaller fragments to move faster than larger fragments.
Transfer to Membrane: The DNA fragments are transferred from the gel onto a solid membrane, typically a positively charged nylon membrane, using capillary action.
Hybridization: The membrane is then exposed to a DNA probe labeled with a radioactive, fluorescent, or chemical tag. The probe is designed to be complementary to the target DNA sequence, allowing it to bind to the specific DNA fragment on the membrane.
Detection: The bound probe is detected using methods such as X-ray film, phosphorimaging, or chemiluminescent substrates, depending on the type of label used.
Southern blotting is used in various applications, including:
Identifying specific DNA sequences in DNA samples
Studying gene rearrangements and mutations
Analyzing viral and bacterial infections
Forensic analysis and personal identification
Gene mapping and restriction enzyme mapping
Identifying methylated sites in genes.
This technique is named after its inventor, Dr. Edwin Southern, who first published it in 1975.
This is all about southern blotting
Blotting
A blot, in molecular biology and genetics, is a method of transferring proteins, DNA or RNA, onto a carrier.
The term "blotting" refers to the transfer of biological samples from a gel to a membrane and their subsequent detection on the surface of the membrane.
Types of blotting techniques
Southern Blotting
Northern Blotting
Western Blotting
A Southern blot is a method used
in molecular biology for detection of a specific DNA sequence in DNA samples.
Southern blotting combines transfer of electrophoresis -separated DNA fragments to a filter membrane and subsequent fragment detection by probe hybridization.
The method is named after its inventor, the British biologist Edwin Mellor Southern.
- Methods in Southern blotting
- Advantages and disadvantages
These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
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.
Couples presenting to the infertility clinic- Do they really have infertility...Sujoy Dasgupta
Dr Sujoy Dasgupta presented the study on "Couples presenting to the infertility clinic- Do they really have infertility? – The unexplored stories of non-consummation" in the 13th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2024) at Manila on 24 May, 2024.
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.
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.
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
These lecture slides, by Dr Sidra Arshad, offer a quick overview of physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar leads (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
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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.
2. 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.
3. TYPES OF BLOTTING TECHNIQUES
Blotting technique
It is used to detect protein.
Western blotNorthern Blot
It is used to detect RNA.
It is used to detect DNA.
Southern Blot
4. Creating the Sandwich
The sandwich consists of :
filter paper
Nitrocellulose membrane
gel matrix
another piece of filter paper
5. Cont….
• First of all two pieces of filter paper and a piece
of nitrocellulose membrane to an appropriate
size is taken and soak them in transfer buffer.
• A piece of buffer soaked filter paper is placed
over the gel.
• The buffer soaked nitrocellulose membrane is
placed against the exposed gel.
• The second piece of buffer soaked filter paper is
placed on the nitrocellulose membrane (covering
it)
• There should not be bubbles between the
membrane and the gel.
6. SOUTHERN BLOTTING
• This method Involves separation,transfer
and hybridization
• The Southern blot is used to detect the presence of a
particular piece of DNA in a sample.
• The DNA detected can be a single gene,or it can be part
of a larger piece of DNA such as a viral genome
• 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
7. 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.
8. Steps in southern blotting Technique
Genomic DNA
Restriction endonuclease
DNA fragments
Gel electrophoresis
Long DNA fragments and short DNA fragments
on Agarose gel
Denatured by mild alkali blot transfer
Nitrocellulose or (nylon membrane)
10. APPLICATIONS
• Southern blots are used in gene discovery ,mapping, evolution
and development studies,diagnostics and forensics.
• In regards to genetically modified organisms,Southern blotting
is used for testing to ensure that a particular section of DNA
of known genetic sequence has been successfully
incorporated into the genome of the host organism.
•Southern blots allow investigators to determine the molecular
weight of a restriction fragment and to measure relative
amounts in different samples.
11. CONT…
•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.
•Analyze restriction digestion fragmentation of
DNA or a biological sample
12. 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 and was named such by
analogy to Southern blotting.
13. An outline of Northern blotting
RNA extract
Agarose gel electrophoresis
rRNA bands
Blotting Hybridization Autoradiography
DNA probe hybridizes to RNA
15. APPLICATIONS
• A standard for the direct study of gene expression at the level of mRNA
(messenger RNA transcripts).
• Detection of mRNA transcript size
• Study RNA degradation
• Study RNA splicing - can detect alternatively spliced transcripts
• Study RNA half-life
• Study IRES (internal ribosomal entry site) – to remove possibility
of RNA digestion .
• Often used to confirm and check transgenic.
16. Disadvantage of Northern blotting
•Often radioactivity is used. This prevents ease of
performing it, use and disposal. New methods of non-
radioactive detection have been generated allowing non-
radioactive detection.
•The whole process of northern blotting takes along time
usually, from sample preparation through to detection.
•If RNA samples are even slightly degraded by RNases,
the quality of the data and quantitation of expression is
quite negatively affected.
17. Western blotting
•Western blotting is an Immunoblotting technique
which rely on the specificity of binding between a
molecule of interest and a probe to allow detection
of the molecule of interest in a mixture of many
other similar molecules.
•In Western blotting, the molecule of interest is a
protein and the probe is typically an antibody raised
against that particular protein.
•The SDS PAGE technique is a prerequisite for
Western blotting
18. Steps in western blotting
• A protein sample is subjected to
electrophoresis on an SDS-polyacrylamide
gel.
•Electroblotting transfers the separated
proteins from the gel to the surface of a
nitrocellulose membrane.
19. CONT….
• The blot is incubated with a generic protein (such
as milk proteins or BSA) which binds to any
remaining sticky places on the nitrocellulose.
• 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,forming a layer
of antibody molecules .
20.
21. Applications
• The confirmatory HIV test employs a Western blot to detect anti-
HIV antibody in a human serum sample. Proteins from known
HIV-infected cells are separated and blotted on a membrane
then,the serum to be tested is applied in the primary antibody
incubation step; free antibody is washed away, and a secondary
anti-human antibody linked to an enzyme signal is added. The
stained bands then indicate the proteins to which the patient's
serum contains antibody.
• A Western blot is also used as the definitive test for Bovine
spongiform encephalopathy (BSE,commonly referred to as 'mad
cow disease').
• Some forms of Lyme disease testing employ Western blotting.