Origin and Characteristics of the cell line: 3D7 Origin: Cloned from NF54. Characteristics: Fully sequenced genome (reference strain for P. falciparum genome projects). Drug-sensitive (wild-type for pfcrt and pfmdr1). High transfection efficiency (commonly used in molecular genetics studies). Dd2 Origin: Isolated from a patient in Indochina (Cambodia). Characteristics: Multidrug-resistant (resistant to chloroquine, mefloquine, and quinine). Key mutations: pfcrt (K76T) → Chloroquine resistance pfmdr1 amplification → Mefloquine resistance Lower transfection efficiency than 3D7. Applications: Drug resistance studies, genetic variation analysis.

CRISPR became widely available as a gene-editing tool in **2012-2013** after key scientific breakthroughs. - 2012: Jennifer Doudna and Emmanuelle Charpentier** published a landmark paper in *Science* demonstrating that the CRISPR-Cas9 system could be programmed to cut specific DNA sequences in vitro (**Jinek et al., 2012**). - 2013: Feng Zhang, George Church, and colleagues adapted CRISPR-Cas9 for genome editing in **mammalian cells**, proving its potential as a gene-editing tool (**Cong et al., 2013; Mali et al., 2013**). By 2014-2015, CRISPR was widely adopted in molecular biology laboratories for gene editing across various organisms, including plants, animals, and human cells.

Thank you for sharing the information. Very educative

Thanks for sharing!

hi thanks for this video! what if I don't know the effect size and I can't calculate de mean or SD, ( I don't have any data and Im working with a novel animal model) ? I have used the number 0.5 for effect size in different calculators like clincalc and kingdom but they give me different numbers for sample size! what can I do? which one should I use.

Hello mam thanks for this life saving video .i am a first year masters student in china studying molecular biology.Mam, i will really need your advise and help in some areas how can i get in touch with you please ma??

Thank you!!

Thanks, howevr, the formula used here is :C1V1= C2V2, Where C1 =50mg/ml, V1=to find, C2=0.4mg/ml, V2=200. Cheers:)

You could also mention that log 5 / log 2 is actually equal to log2 (5) = c. 2.3219 - a little bit more than 2 which is log2(4}. 😊

thank you

📖 Additional Reading: Read and Hyde, Simple In Vitro Cultivation of the Malaria Parasite: bit.ly/3D1eyvt Adwoa Biotech Blog: bit.ly/41gtxvu

Blood groups: kzbin.info/www/bejne/mau5mKWobZZnpqcsi=HOOXGG0VRHk8M7aB

Which strain of Plasmodium falciparum parasite do you have in your lab? Here's the video on the parasite life stages: kzbin.info/www/bejne/nHvMknSJhax2rZosi=UEzIA9IQs0-GcrIX

Thanks

Keep going sis one day you'll achieve 1million subscriber 0:46

Reagents for Calcium Phosphate: Plasmid DNA Transfection: 1) Calcium chloride (CaCl₂), 2.5 M 2) 2× HEPES-buffered saline (HBS): (280 mM NaCl, 50 mM HEPES (pH 7.1), 1.5 mM Na₂HPO₄). Filter sterilize and store at 4°C 3) Plasmid DNA (0.5 to 10 µg)

Thank you so much for this video!

➡1880 Charles Louis Alphonse Laveran identifies the malaria parasite. 1898 Sir Ronald Ross demonstrates that mosquitoes transmit malaria. 1976 William Trager and JB Jensen grow parasite in culture for first time.

Anytime you need to copy DNA, whether it’s for diagnostic assays, forensic analysis or gene cloning, you will need to design primers. PCR or cloning primers guide polymerases to amplify gene-specific sequences. They are designed to flank the target region (region to be copied), to allow DNA synthesis and replication to occur.

Timeline of progress fighting malaria (Adapted from www.ucsf.edu/news/2011/08/103884/progress-fighting-malaria-timeline): 1820 Quinine first purified from tree bark. For many years prior, the ground bark had been used to treat malaria. 1934 Hans Andersag in Germany discovers the Anti-malarial drug Chloroquine, which is not widely used until after World War II. 1957 First documented case of resistance to Chloroquine is reported. 1989 The U.S. Food and Drug Administration approves the use of the anti-malaria drug Mefloquine hydrochloride, registered as Lariam®by Hoffman-LaRoche. 1992 Malaria vaccine candidate RTS,S, developed by GlaxoSmithKline and the Walter Reed Army Institute of Research, enters clinical trials. 1996 Insecticide-treated bednets are proven to reduce overall childhood mortality by 20 percent in large, multi-country African study. 2001 WHO prequalifies first fixed-dose Artemisinin combination therapy (ACT), sold by Novartis as Coartem® and recommends ACT as first-line malaria treatment. 2002 Genome sequencing of Anopheles gambiae (mosquito) and Plasmodium falciparum (parasite) completed. 2007 UCSF study shows combination malaria therapy effective in treating African children 2008 Rectal application of the inexpensive antimalarial drug artesunate proven to save the lives of young children with severe malaria.

When William Trager and JB Jensen grew plasmodium parasites in culture for the first time in 1976, it opened the way for drug discovery and vaccine research.

Timeline of progress fighting malaria: www.ucsf.edu/news/2011/08/103884/progress-fighting-malaria-timeline

If I'm starting with blood that has had the plasma removed do I add PBS in a ratio of 1:1 to the original amount of blood (PRIOR) to the plasma removed or 1:1 to the volume of blood remaining (AFTER) the plasma is removed?

Western blotting is a technique used to detect specific proteins in a sample. It involves the following key steps: - Sample Preparation: Proteins are extracted from biological samples and denatured. - Gel Electrophoresis: Proteins are separated by size using SDS-PAGE. - Transfer: Proteins are transferred to a membrane (usually nitrocellulose or PVDF). - Blocking: Non-specific binding sites on the membrane are blocked. - Antibody Incubation: The membrane is incubated with primary and secondary antibodies. - Detection: The presence of the target protein is visualised.

Western blotting is a technique used to detect specific proteins in a sample. It involves the following steps: - Sample Preparation: where proteins are extracted from biological samples and denatured. - Gel Electrophoresis: where proteins are separated by size using Sodium Dodecyl Sulfate (SDS)-PAGE. - Transfer: where proteins are transferred to a membrane (usually nitrocellulose or PVDF). - Blocking: where non-specific binding sites on the membrane are blocked. - Antibody Incubation: The membrane is incubated with primary and secondary antibodies. - Detection: The presence of the target protein is visualized House keeping genes are used to demonstrate that there no variations to the proteins occurred, during any of the steps.

W!!!

Nice video..

An easier way to describe this would be to simply say a 100% solution is 1 gram of solute in 1 ml of solvent

becuz scratch

Thanks

Hi, when doing gRNA design for HDR , should one choose Knockin or Knockout option from choochop software. The intention is to provide repair temple during KOs

Bauch Drives

False about Jefferson, Jefferson was an actual great abolitionist, if he actually did make a statement, then I am sure it is taken out of context, like many of the founders words and actions

ADDITIONAL VALIDATION APPROACHES The fusion of eGFP to a protein additionally allows researchers to observe the spatial and temporal localization of that protein inside live cells using fluorescence microscopy. eGFP (enhanced Green Fluorescent Protein) can also be used to track and compare protein-protein interactions in wild-type (WT) and mutant proteins through immunoprecipitation (IP) experiments. Immunoprecipitation allows the isolation of a protein of interest from a cell lysate, often along with its interacting partners. Proteins can also be tagged with small epitope tags such as FLAG, HA, and Myc during expression to facilitate purification, detection, and analysis.

Yes, quality makes a lab more confident.

well explained

Have you been prescribed any of these antimalarials before?

Great video!! Glad our paths crossed!

I have to add a comment. THANK YOU! I am an attorney and I am taking a class in cybersecurity that is way more mathy than my law school-trained brain is accustomed to. This video is easy to follow and very helpful. I am glad you gave us a couple of examples to practice and also, the "why" of using the log function was interesting. I do not remember much from my trig days in high school, many years ago. Again, thank you!

Thank you Adwoa for making this video. Most of these content are put together by the Indians and westerners. I’m glad you attempted this. I know you will serve us with even better content. You did well

I should become a doctor in my next life ❤

Thanks Adwoa! We need more teachers like you to help us understand scientific concepts better! Great tutorial

Hi, great video. I am attempting to do this for SDS-PAGE gels rather than WB ones. I am trying to compare Rubisco content and purity from various treatments. However, unlike WB where you probe for a target protein (e.g Rubisco), and get one nice band, my SDS-PAGE gel has many bands other than my POI. In this case, where would I subtract the blank from as there are no 'empty' spaces in the lanes? I thought of taking three measurements at the top of the gel (not in the lanes) and taking an average of this as the background. Do you have any suggestions? Also, I am not sure about the case where there are multiple bands per lane and the fact that I want to find out relative purity and compare the relative quantity between treatments (using a standard curve generated from Rubisco quantitation standard) if I should use the integrated density method or the NIH (AUC) method? Please give me any tips and suggestions. Thanks!

Original Paper Electrophoretic Transfer of Proteins from Polyacrylamide Gels to Nitrocellulose Sheets: Procedure and Some Applications: www.pnas.org/content/pnas/76/9/4350.full.pdf

References to deepen your knowledge : Books "Principles of Instrumental Analysis" by Douglas A. Skoog, F. James Holler, Stanley R. Crouch This book provides comprehensive coverage of analytical techniques, including the use of internal standards in various instrumental methods. ISBN: 978-0495012016 "Quantitative Chemical Analysis" by Daniel C. Harris This text covers the principles and practices of quantitative analysis, including the application of internal standards in different analytical procedures. ISBN: 978-1464135385 "Handbook of Analytical Techniques" edited by Helmut Günzler and Alex Williams This handbook includes detailed discussions on various analytical techniques and the use of internal standards to improve data quality. ISBN: 978-3527296539 Articles and Papers "Use of Internal Standards in Quantitative Analytical Chemistry" by Alan G. Marshall and Bruce S. Meyer Journal of Chemical Education, 1977, Vol. 54, No. 10, pp. 640-643. This article provides an overview of the principles and applications of internal standards in quantitative analysis. DOI: 10.1021/ed054p640 "The Role of Internal Standards in Quantitative Analysis by Mass Spectrometry" by Chris B. Stack, David Muddiman Analytical Chemistry, 2011, Vol. 83, No. 12, pp. 4649-4660. Discusses the use of internal standards in mass spectrometry to enhance quantitative accuracy. DOI: 10.1021/ac200160w "Application of Internal Standards in Chromatographic Analysis" by James W. Taylor Journal of Chromatographic Science, 2002, Vol. 40, No. 6, pp. 292-299. Explores the use of internal standards in chromatographic methods to improve quantification. DOI: 10.1093/chromsci/40.6.292 Online Resources National Institute of Standards and Technology (NIST) - Analytical Chemistry Division The NIST website provides guidelines and resources related to analytical methods, including the use of internal standards. NIST Analytical Chemistry Division International Union of Pure and Applied Chemistry (IUPAC) - Compendium of Chemical Terminology (Gold Book) The IUPAC Gold Book offers definitions and explanations of terms used in analytical chemistry, including internal standards. IUPAC Gold Book Practical Guides "Quantitative Analysis Using Internal Standards in HPLC" by Jürgen Hochstrasser A practical guide on the application of internal standards in high-performance liquid chromatography (HPLC). Available online as a technical note from various chromatography companies.

Hi! What temperature do you usually spin the sample at? Room? 4ºC? Thank you so much!

The teaching methodology is not good...

bullshit, not helpfull at all

I did DNA extraction by method of Maxi prep for the first time. I didn’t even understand the others steps. I’m willing to learn more methods and other steps.

Greetings from America. I love your teaching approach Adwoa. Your channel is one of my favorites! Can you create a video similar to this showing how internal standards are used for a calibration curve. From what I understand ratios are key for the internal standard approach. Any information on this would be much appreciated!