This presentation contains information about restriction enzymes, its nomenclature, restriction digestion, and its application. This also contains information about the chemicals used in restriction and also explains the general procedure of restriction digestion of DNA
This presentation is prepared to provide a general overview of Recombinant DNA technology.
The flow of the presentation is following manner.
Slide 1. Introduction
Slide 2. The basic principle of Recombinant DNA technology
Slide 3. Restriction endonucleases
Slide 4. Cloning
Slide 5. Vectors
Slide 6. Transformation
Slide 7. Screening of clones
Slide 8. Sequencing and polymerase chain reaction
Slide 9. Case Study: Disease identification and therapy discovery
This presentation contains information about restriction enzymes, its nomenclature, restriction digestion, and its application. This also contains information about the chemicals used in restriction and also explains the general procedure of restriction digestion of DNA
This presentation is prepared to provide a general overview of Recombinant DNA technology.
The flow of the presentation is following manner.
Slide 1. Introduction
Slide 2. The basic principle of Recombinant DNA technology
Slide 3. Restriction endonucleases
Slide 4. Cloning
Slide 5. Vectors
Slide 6. Transformation
Slide 7. Screening of clones
Slide 8. Sequencing and polymerase chain reaction
Slide 9. Case Study: Disease identification and therapy discovery
BRIEFLY EXPLAINED PPT ABOUT RESTRICTION ENZYMES, THEIR WORKING SITES, TYPES, ARTIFICIALLY GENERATED RESTRICTION ENZYMES, THEIR MECHANISM OF ACTION, TYPES OF CUTS THEY MAKE, THEIR NOMENCLATURE ETC.
Plasmid is a double stranded, circular extra chromosomal DNA of bacterium. It is used in recombinant DNA experiments to clone genes from other organisms and make large quantities of their DNA. Plasmid can be transferred between same species or between different species. Size of plasmids range from 1-1000 kilo base pairs. Plasmids are part of mobilomes (total of all mobile genetic elements in a genome) like transposons or prophages and are associated with conjugation. Even the largest plasmids are considerably smaller than the chromosomal DNA of the bacterium, which can contain several million base pairs.
Restriction Endonuclease: The Molecular Scissor of DNA - By RIKI NATHRIKI NATH
restriction enducleases are called the molecular scissors of DNA. types of restriction enzymes, their structures, subunits, most importantly the use of Type II restriction endonuclease in recombinant technology, mechanism of enzyme action and their applications.
DNA consists of a linear string of nucleotides, or bases, for simplicity, referred to by the first letters of their chemical names--A, T, C and G. The process of deducing the order of nucleotides in DNA is called DNA sequencing. Since the DNA sequence confers information that the cell uses to make RNA molecules and proteins, establishing the sequence of DNA is key for understanding how genomes work. The technology for DNA sequencing was made faster and less expensive as a part of the Human Genome Project. And recent developments have profoundly increased the efficiency of DNA sequencing even further.
BRIEFLY EXPLAINED PPT ABOUT RESTRICTION ENZYMES, THEIR WORKING SITES, TYPES, ARTIFICIALLY GENERATED RESTRICTION ENZYMES, THEIR MECHANISM OF ACTION, TYPES OF CUTS THEY MAKE, THEIR NOMENCLATURE ETC.
Plasmid is a double stranded, circular extra chromosomal DNA of bacterium. It is used in recombinant DNA experiments to clone genes from other organisms and make large quantities of their DNA. Plasmid can be transferred between same species or between different species. Size of plasmids range from 1-1000 kilo base pairs. Plasmids are part of mobilomes (total of all mobile genetic elements in a genome) like transposons or prophages and are associated with conjugation. Even the largest plasmids are considerably smaller than the chromosomal DNA of the bacterium, which can contain several million base pairs.
Restriction Endonuclease: The Molecular Scissor of DNA - By RIKI NATHRIKI NATH
restriction enducleases are called the molecular scissors of DNA. types of restriction enzymes, their structures, subunits, most importantly the use of Type II restriction endonuclease in recombinant technology, mechanism of enzyme action and their applications.
DNA consists of a linear string of nucleotides, or bases, for simplicity, referred to by the first letters of their chemical names--A, T, C and G. The process of deducing the order of nucleotides in DNA is called DNA sequencing. Since the DNA sequence confers information that the cell uses to make RNA molecules and proteins, establishing the sequence of DNA is key for understanding how genomes work. The technology for DNA sequencing was made faster and less expensive as a part of the Human Genome Project. And recent developments have profoundly increased the efficiency of DNA sequencing even further.
Blotting technique including Southern , Northern and Western blotting Rohit Mondal
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Class9 DNA technology in secondary schoolssusera700ad
Biotechnology is the use of an organism, or a component of an organism or other biological system, to make a product or process.
Many forms of modern biotechnology rely on DNA technology.
DNA technology is the sequencing, analysis, and cutting-and-pasting of DNA.
Common forms of DNA technology include DNA sequencing, polymerase chain reaction, DNA cloning, and gel electrophoresis.
Biotechnology inventions can raise new practical concerns and ethical questions that must be addressed with informed input from all of society.
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Length: 30 minutes
Session Overview
-------------------------------------------
During this webinar, we will cover the following topics while demonstrating the integrations of JMeter, InfluxDB and Grafana:
- What out-of-the-box solutions are available for real-time monitoring JMeter tests?
- What are the benefits of integrating InfluxDB and Grafana into the load testing stack?
- Which features are provided by Grafana?
- Demonstration of InfluxDB and Grafana using a practice web application
To view the webinar recording, go to:
https://www.rttsweb.com/jmeter-integration-webinar
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Interested in deploying notification automations for Bonterra Impact Management? Contact us at sales@sidekicksolutionsllc.com to discuss next steps.
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Keynote at DIGIT West Expo, Glasgow on 29 May 2024.
Cheryl Hung, ochery.com
Sr Director, Infrastructure Ecosystem, Arm.
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- Visualization tools to display your network;
- Grid simulation tools, such as power flows, security analyses (with or without remedial actions) and sensitivity analyses;
The framework is mostly written in Java, with a Python binding so that Python developers can access PowSyBl functionalities as well.
What you will learn during the webinar:
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UI automation Sample
Desktop automation flow
Pradeep Chinnala, Senior Consultant Automation Developer @WonderBotz and UiPath MVP
Deepak Rai, Automation Practice Lead, Boundaryless Group and UiPath MVP
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• Test Automation: How AI-powered test case generation, optimization, and self-healing tests are making testing more efficient and effective.
• Visual Testing: Explore the emerging capabilities of AI in visual testing and how it's set to revolutionize UI verification.
• Inflectra's AI Solutions: See demonstrations of Inflectra's cutting-edge AI tools like the ChatGPT plugin and Azure Open AI platform, designed to streamline your testing process.
Whether you're a developer, tester, or QA professional, this webinar will give you valuable insights into how AI is shaping the future of software delivery.
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Transcript: Selling digital books in 2024: Insights from industry leaders - T...BookNet Canada
The publishing industry has been selling digital audiobooks and ebooks for over a decade and has found its groove. What’s changed? What has stayed the same? Where do we go from here? Join a group of leading sales peers from across the industry for a conversation about the lessons learned since the popularization of digital books, best practices, digital book supply chain management, and more.
Link to video recording: https://bnctechforum.ca/sessions/selling-digital-books-in-2024-insights-from-industry-leaders/
Presented by BookNet Canada on May 28, 2024, with support from the Department of Canadian Heritage.
8. SCREENING OF cdna library...
The result of any cloning experiment that
begins with total DNA from specific source is a
library of clones.
One of the key elements required to
Identify the gene during cloning is known
As Probe.
A Probe is normally a cloned piece of
DNA that contains a portion of the sequence
For which you are searching.
Typically , will make the probe radioactive
And add it to a solution.
This is binding step is known as
Hybridisation.
9.
10.
11. Types of screening methods:
By colony hybridisation,
By plaque hybridisation,
With labelled oligonucleotide probe,
Immunological screening,
Differential screening,
Blue-white screening,
By replica plating,
Expression screening.
12. Colony hybridisation...
• Colony hybridisation is the “BLOT ANALYSIS
TECHNIQUE” where the bacterial cells are transferred
From the solid nutrient medium to the absorbent
Material.
• It can defines as a method for the isolation of
The specific DNA sequence or genes from the
Bacterial cells containing hybrid DNA, by the
Means of a nitrocellulose membrane filter.
• The transferring medium then goes through
Several chemicals and physical treatment.
13. Colony hybridisation involves the
following steps:
First, inoculate the bacterial cell suspension o the
Solid agar medium to prepare the master plate.
After inoculation, the number of bacterial colonies
Will develope with different plasmids which refer
As “master or reference plate”.
Then transfer the bacterial cells from the master
Plate on to the membrane or filter by the means of
Nitrocellulose filter.
Press the nitrocellulose paper over the surface of
The master plate.
14. Cont.....
The compression of the filter membrane will form
Replicas or copies of the bacterial cella as that of master
plate.
Then the treatment of filter medium with SDS to
lyse the bacterial cells.
Treat the filter paper with alkali to separate the
DNA into single strands & fixation of dna onto the
filter medium.
Addition of radioactive probe; it will code the
desired gene of sequence from the bacterial cells&then
washing and autoradiography.
15.
16. Plaque hybridisation...
Plaque hybridisation is a technique use in
molecular biology for the identification of recombinant
phages.
The procedure can also be used for the detection
of differently represented repetitive dna.
The technique [similar to colony hybridisation]
involves hybridising isolated phage DNA to a
label probe for the gene of study.
The plaque hybridisation procedure has some
advantages over colony hybridisation due to the
smaller and well defined area of the filter to which
the DNA binds.
18. With labelled oligoucleotide probe:
This type of screening based on a known
peptide sequence.
The protein encoded by the gene of interest is
first digest with protease to breakdown as a amino
acids, and separate the each peptides.
Then treat the peptides with the Edman’s
Reagent.
The target sequence are complimentary
sequence of DNA which is used as a primer.
19.
20. Immunological screening...
Immunological screening of an expression c DNA
primary antibody and labelled secondary antibody;
note the label is often an enzyme label like alkaline
phosphate ,but it also be I125 .
21.
22. Differential screening:
o Differential screening is probably the most
Direct approach for the identification of new genes
Whose expression is associated with a change in
Physiological conditions.
o In the recent years , other approaches have
Been developed, including the production of
Of substracted Cdna libraries , differential
Display and RNA fingerprinting.
23.
24. Blue-white screening...
It is mostly used in recombinant DNA
technology, sometimes this method is used in
screening of c DNA library.
Foreign DNA is inserted into the plasmid,
where it inactivates the lac z gene.
The recombinant plasmid is introduced
Into a bacterium which become amp-resistant
All treated bacteria are spread on a nutrient
Agar plate containing ampicillin and beta-
galactosidase substrate and incubated.
When colonies that appear must contain foreign
DNA .
Blue colonies do not contain foreign DNA.
25.
26. Replica-plating...
• Replica plating is a microbiological technique
In which one or more secondary Petri plates
Containing different solid selective growth media
Are inoculated with the same colonies of
Micro organisms from a primary plate,
Reproducing the original spatial pattern of
Colonies.
• The technique involves pressing a velveteen
Covered disk , and then imprinting secondary
Plates with cells in colonies removed from the
Original plate by the material.
• Replica plating is especially useful for negative
Selection.
27. • For example, if one want to select the colonies that
Were sensitive to AMPICILLIN , the plate could be
Replica plated on a secondary ampicillin agar plate.
• The purpose of replica plating is to be able to compare
The master plate and secondary plates, typically to
Screen for a desired phenotype.
• Common screenable phenotypes include
auxotrophy and antibiotic resistance.
28.
29. Expression screening...
Antibodies used to screen the expression library.
The procedure has similarities to the plaque
hybridisation protocol.
plaque lift (taking by placing a
membrane on the dish of the plaque)
immersed in a solution of the
antibody.
detected by other antibodies.
repeat cycles of screening to
isolate pure plaques.