This is a powerpoint file of a practical class taken by Dr. Karthikeyan Pethsuamay for the first year MBBS students of AIIMS, New Delhi. Feel free to download and use for educational purposes. Happy learning and teaching!
Don't forget to watch the YouTube video.
This is a powerpoint file of a practical class taken by Dr. Karthikeyan Pethsuamay for the first year MBBS students of AIIMS, New Delhi. Feel free to download and use for educational purposes. Happy learning and teaching!
Don't forget to watch the YouTube video.
Reverse transcription of RNA is a process whereby RNA, typically messenger RNA is converted into complimentary DNA. The process was discovered by Howard Temin and John Baltimore when they observed that certain RNA viruses could revert to DNA. This was an important discovery that led to the discovery of enzymes classified as reverse transcriptases. Today Reverse Transcription is routinely applied in molecular biology laboratories to obtain the stable cDNA version of RNA for downstream analysis.
Polymerase chain reaction (PCR) is a technique used in molecular biology to amplify a single copy or a few copies of a segment of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. It is an easy, cheap, and reliable way to repeatedly replicate a focused segment of DNA, a concept which is applicable to numerous fields in modern biology and related sciences.
A biochemical technique used in Molecular Biology to amplify a specific fragment of target DNA.
PCR is used in medical and biological research, including cloning, genetic analysis, genetic fingerprinting, diagnostics, pathogen detection and genetic fingerprinting
Polymerase chain reaction or PCR is a laboratory technique that has been elaborated in many ways since its introduction and is now commonly used for a wide variety of applications including genotyping, cloning, mutation detection, sequencing, microarrays, forensics, and paternity testing.
A detailed description about the basic steps involved in the - PCR - Polymerase Chain Reaction, its applications,its limitations and steps to overcome it.
Reverse transcription of RNA is a process whereby RNA, typically messenger RNA is converted into complimentary DNA. The process was discovered by Howard Temin and John Baltimore when they observed that certain RNA viruses could revert to DNA. This was an important discovery that led to the discovery of enzymes classified as reverse transcriptases. Today Reverse Transcription is routinely applied in molecular biology laboratories to obtain the stable cDNA version of RNA for downstream analysis.
Polymerase chain reaction (PCR) is a technique used in molecular biology to amplify a single copy or a few copies of a segment of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. It is an easy, cheap, and reliable way to repeatedly replicate a focused segment of DNA, a concept which is applicable to numerous fields in modern biology and related sciences.
A biochemical technique used in Molecular Biology to amplify a specific fragment of target DNA.
PCR is used in medical and biological research, including cloning, genetic analysis, genetic fingerprinting, diagnostics, pathogen detection and genetic fingerprinting
Polymerase chain reaction or PCR is a laboratory technique that has been elaborated in many ways since its introduction and is now commonly used for a wide variety of applications including genotyping, cloning, mutation detection, sequencing, microarrays, forensics, and paternity testing.
A detailed description about the basic steps involved in the - PCR - Polymerase Chain Reaction, its applications,its limitations and steps to overcome it.
This presentation was given in 2009 at Insigma HengTian in Hangzhou, China as an overview of the causes of the Financial Crisis. The goals of this presentation were to clarity commonly used English vocabulary and explain differences between Chinese and American financial structures.
JMeter webinar - integration with InfluxDB and GrafanaRTTS
Watch this recorded webinar about real-time monitoring of application performance. See how to integrate Apache JMeter, the open-source leader in performance testing, with InfluxDB, the open-source time-series database, and Grafana, the open-source analytics and visualization application.
In this webinar, we will review the benefits of leveraging InfluxDB and Grafana when executing load tests and demonstrate how these tools are used to visualize performance metrics.
Length: 30 minutes
Session Overview
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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
Encryption in Microsoft 365 - ExpertsLive Netherlands 2024Albert Hoitingh
In this session I delve into the encryption technology used in Microsoft 365 and Microsoft Purview. Including the concepts of Customer Key and Double Key Encryption.
UiPath Test Automation using UiPath Test Suite series, part 3DianaGray10
Welcome to UiPath Test Automation using UiPath Test Suite series part 3. In this session, we will cover desktop automation along with UI automation.
Topics covered:
UI automation Introduction,
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
Software Delivery At the Speed of AI: Inflectra Invests In AI-Powered QualityInflectra
In this insightful webinar, Inflectra explores how artificial intelligence (AI) is transforming software development and testing. Discover how AI-powered tools are revolutionizing every stage of the software development lifecycle (SDLC), from design and prototyping to testing, deployment, and monitoring.
Learn about:
• The Future of Testing: How AI is shifting testing towards verification, analysis, and higher-level skills, while reducing repetitive tasks.
• 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.
LF Energy Webinar: Electrical Grid Modelling and Simulation Through PowSyBl -...DanBrown980551
Do you want to learn how to model and simulate an electrical network from scratch in under an hour?
Then welcome to this PowSyBl workshop, hosted by Rte, the French Transmission System Operator (TSO)!
During the webinar, you will discover the PowSyBl ecosystem as well as handle and study an electrical network through an interactive Python notebook.
PowSyBl is an open source project hosted by LF Energy, which offers a comprehensive set of features for electrical grid modelling and simulation. Among other advanced features, PowSyBl provides:
- A fully editable and extendable library for grid component modelling;
- 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:
- For beginners: discover PowSyBl's functionalities through a quick general presentation and the notebook, without needing any expert coding skills;
- For advanced developers: master the skills to efficiently apply PowSyBl functionalities to your real-world scenarios.
Securing your Kubernetes cluster_ a step-by-step guide to success !KatiaHIMEUR1
Today, after several years of existence, an extremely active community and an ultra-dynamic ecosystem, Kubernetes has established itself as the de facto standard in container orchestration. Thanks to a wide range of managed services, it has never been so easy to set up a ready-to-use Kubernetes cluster.
However, this ease of use means that the subject of security in Kubernetes is often left for later, or even neglected. This exposes companies to significant risks.
In this talk, I'll show you step-by-step how to secure your Kubernetes cluster for greater peace of mind and reliability.
Generating a custom Ruby SDK for your web service or Rails API using Smithyg2nightmarescribd
Have you ever wanted a Ruby client API to communicate with your web service? Smithy is a protocol-agnostic language for defining services and SDKs. Smithy Ruby is an implementation of Smithy that generates a Ruby SDK using a Smithy model. In this talk, we will explore Smithy and Smithy Ruby to learn how to generate custom feature-rich SDKs that can communicate with any web service, such as a Rails JSON API.
State of ICS and IoT Cyber Threat Landscape Report 2024 previewPrayukth K V
The IoT and OT threat landscape report has been prepared by the Threat Research Team at Sectrio using data from Sectrio, cyber threat intelligence farming facilities spread across over 85 cities around the world. In addition, Sectrio also runs AI-based advanced threat and payload engagement facilities that serve as sinks to attract and engage sophisticated threat actors, and newer malware including new variants and latent threats that are at an earlier stage of development.
The latest edition of the OT/ICS and IoT security Threat Landscape Report 2024 also covers:
State of global ICS asset and network exposure
Sectoral targets and attacks as well as the cost of ransom
Global APT activity, AI usage, actor and tactic profiles, and implications
Rise in volumes of AI-powered cyberattacks
Major cyber events in 2024
Malware and malicious payload trends
Cyberattack types and targets
Vulnerability exploit attempts on CVEs
Attacks on counties – USA
Expansion of bot farms – how, where, and why
In-depth analysis of the cyber threat landscape across North America, South America, Europe, APAC, and the Middle East
Why are attacks on smart factories rising?
Cyber risk predictions
Axis of attacks – Europe
Systemic attacks in the Middle East
Download the full report from here:
https://sectrio.com/resources/ot-threat-landscape-reports/sectrio-releases-ot-ics-and-iot-security-threat-landscape-report-2024/
Kubernetes & AI - Beauty and the Beast !?! @KCD Istanbul 2024Tobias Schneck
As AI technology is pushing into IT I was wondering myself, as an “infrastructure container kubernetes guy”, how get this fancy AI technology get managed from an infrastructure operational view? Is it possible to apply our lovely cloud native principals as well? What benefit’s both technologies could bring to each other?
Let me take this questions and provide you a short journey through existing deployment models and use cases for AI software. On practical examples, we discuss what cloud/on-premise strategy we may need for applying it to our own infrastructure to get it to work from an enterprise perspective. I want to give an overview about infrastructure requirements and technologies, what could be beneficial or limiting your AI use cases in an enterprise environment. An interactive Demo will give you some insides, what approaches I got already working for real.
Smart TV Buyer Insights Survey 2024 by 91mobiles.pdf91mobiles
91mobiles recently conducted a Smart TV Buyer Insights Survey in which we asked over 3,000 respondents about the TV they own, aspects they look at on a new TV, and their TV buying preferences.
Slack (or Teams) Automation for Bonterra Impact Management (fka Social Soluti...Jeffrey Haguewood
Sidekick Solutions uses Bonterra Impact Management (fka Social Solutions Apricot) and automation solutions to integrate data for business workflows.
We believe integration and automation are essential to user experience and the promise of efficient work through technology. Automation is the critical ingredient to realizing that full vision. We develop integration products and services for Bonterra Case Management software to support the deployment of automations for a variety of use cases.
This video focuses on the notifications, alerts, and approval requests using Slack for Bonterra Impact Management. The solutions covered in this webinar can also be deployed for Microsoft Teams.
Interested in deploying notification automations for Bonterra Impact Management? Contact us at sales@sidekicksolutionsllc.com to discuss next steps.
11. Final Heparin Column Final Heparin Column Mark 80 HepFT HepW HepE7 HepE6 HepE5 HepE4 HepE3 HepE2 HepE1
12.
13. ATPase Assay MfdC ATP ADP+P i Pyruvate Kinase Phosphoenolpyruvate Pyruvate Lactate Dehydrogenase Lactate NAD + NADH
14. Activity of MfdC +ATP and DNA Time (minutes) Absorbance at 340nm ATP Avg. = 140 mM ATP/min/ mM enzyme DNA Avg. = 332 mM ATP/min/ mM enzyme - DNA + DNA
15.
Editor's Notes
-Many repair mechanisms work to identify damage of transcribed genes and therefore, genes are much more likely to be repaired after they have been transcribed -Persistence of unrepaired DNA damage can cause mutations and other detrimental factors that can lead to cancer, cell death, aging, and Cockayne’s syndrome -Symptoms of Cockayne’s syndrome impaired nervous system development and premature aging -I am studying the DNA repair protein, Mfd
-Mfd is made up of MfdN and MfdC which are tethered to each other by a linker between them -The are MfdN, which is composed of domains 1 through 3 and MfdC which is composed of domains 4 through 7. MfdC and MfdN are linked together by the linker at domain 3 and domain 4 -I am studying the role of MfdC in transcription coupled DNA repair -MfdC’s role is to bind to DNA during transcription, and to act as an ATP motor for a conformational change
-To understand how Mfd works, I have to give a bit of background on another enzyme, RNA polymerase -When RNA polymerase attempts to transcribe damaged DNA, it will sometimes get stalled and just sit on the DNA -Here is where Mfd comes in -Mfd then recognizes the stalled RNA polymerase, then, the MfdC subunit binds to the DNA behind (upstream of) RNA polymerase. -MfdC uses its ATP motor to cause MfdC and MfdN to open up -The theory is that then MfdN moves to sort of push RNA polymerase off of the DNA -Then, Mfd somehow calls in DNA repair enzymes UvrA, UvrB, nuclease and ligase to repair the damaged DNA
-My goal is to obtain the structure of MfdC when ATP is bound to it and when DNA is bound to it and hopefully gain some insight in the issues above.
-We use a mutant called ATHook-MfdC, which contains a residue that binds well with AT rich areas of DNA -Mike Murphy and Peng Gong found that, compared to Full-Length Mfd, ATHook is superior in terms of activity, binding, and stimulation by DNA
-This is my final purification procedure that we have developed over multiple grow-ups -It’s proven difficult to get MfdC to a pure enough state and at a high enough concentration where we can find consistent crystal results and therefore we need to go through many rigorous steps to get there. -MfdC at high concentrations when expressing in cells can be toxic to the cell, making it a particularly hard protein to express in high amounts.
We did another grow up, this time in 4L to try to get repeatable results in the crystal tray Several samples are missing from the gel we forgot to take some samples along the way The lysate sample precipitated in the loading dye. It was sitting in the dye for a long time and had been cooked twice You can see most of our protein came off in NiE2, that was a 4.5mL fraction Elution buffer is .5M NaCl, 10mM Tris pH 8.2, 250mM Imidazole We then desalted (not shown) and diluted the fractions to 250mM NaCl to load onto the Q column MfdC does not bind to the anion exchange resin so it comes out in the FT FT fractions are from 10-12mL We then eluted contaminant proteins in 4, 2mL fractions designated QE1-QE4 SDS Samples are 7uL
This was a 1mL heparin column It is not very pure, most protein came out in Hep E2 and E3 Each of these are 1mL fractions Hep E2 was taken and loaded onto the sizing column it was at ~2mg/ml SDS Samples are 7uL
Hep E2 was taken from the previous gel and loaded onto the sizing column There was a bubble in the UV detector Each fraction is 3.5mL It came out relatively pure SDS Samples are 7uL
We collected fractions 29-39 from the sizing column, brought them down to 250mM salt and reapplied them to a 1mL heparin column This concentrated our protein (relatively speaking of course) E2 was about 1mg/ml from the Garman lab spec SDS samples are 7uL
Off of our final heparin column we took Hep E2 and buffer exchanged it into .5M Ammonium Acetate; a volatile salt. We then added MgCl2 for a target concentration in the well of 4mM, and AMPpnp for a target concentration in the well of 2mM We used a 1:6 ratio of precipitant to protein The volatile buffer caused the drop to concentrate to 1microliter We got a crystal in C4, which contained .2M Sodium Acetate Trihydrate, .1M Sodium Cacodylate pH 6.5, and 3% w/v PEG 8000 The crystal is about 20micrometers
NADH absorbs light at 340nm Pyruvate Kinase and Lactose dehydrogenase are fast enzymes so we can accurately measure the rate of ATP hydrolysis by measuring the rate of disappearance of NADH
In the past, I have had problems with very low activities, over the summer I figured out that the Ph of our buffers were all off by like 2 ph units. The ATPase assay was done on the DU-800 Spectrophotometer in the Heuck lab at 37 degrees C The activity was measured for 10 minutes, but the assays get wiped out quickly We ignored the nonlinear activity where the substrate concentrations were very low This graph shows how ATHook-MfdC is stimulated twice as much by DNA Concentration of stock protein was about .150mg/mL