We narrowed to 1,475 results for: nin
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TypeBlog Post... feeling of camaraderie one gets from a well-functioning lab team. Obviously not everyone gets this feeling...environments outside of academia provide a formal training regime and the right set of tools to help you ...non-scientists! Maybe you’ve just spent 5-10 years learning to “speak science,” and virtually every person... years of focusing on my technical skills with cloning and lentivirus production, I didn’t really start...
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How to Write a Scientific Review Article
TypeBlog Post...managing a lot of references. I cite as I write, meaning I use the software to add the citations in real...write. Things are going to get a little crazy (meaning you are probably going to cite hundreds of references...better to keep your references organized from the beginning. I also recommend using the citation style of ...relevant to you, note it, and move on. At the beginning I felt like this was cheating. Eventually I learned... -
Antibiotic Resistance: An Old Solution but a New Problem
TypeBlog Post...antibacterial drugs. Resistance can be intrinsic, meaning a basic characteristic that naturally occurs in...especially if the infection is serious or life-threatening, treatment will often begin before this information...results. While this is not the only method of determining drug resistance status, it is a very common, ...essential component of preventing infection, as is cleaning and disinfecting frequently touched surfaces like... -
The Challenges of Cell Culture
TypeBlog Post...immortalized human cell line. He proved this by examining whether the cell lines had isoenzymes and genetic... piece of paper on the front of the incubator containing the passage number, cell line, and date, which...He studied biochemistry at Warwick University, gaining extensive cell culture research experience through...Cell Culture Guide Thermofisher Guidelines for Maintaining Cultured Cells Invitrogen Cell Culture Basics... -
Plasmids 101: Control Plasmids
TypeBlog Post...of control plasmids Part of planning your experiment includes determining what factors need to be controlled...independent variable you are testing and not some other unintended factor. Control plasmids are used to minimize...systems have many variables. However daunting designing and executing these steps may be, proper controls... -
Plasmids 101: E. coli Strains for Protein Expression
TypeBlog Post...of E. coli for DNA propagation. While great for cloning purposes, these E. coli strains are not usually...Use BL21 (DE3) Basic IPTG-inducible strain containing T7 RNAP (DE3) F- ompT lon hsdSB (rB- mB-) gal... (DE3)** Rifampicin RecA-deficient; allows for cloning and expression in same strain F- recA1 hsdR(rK12...most experiments, the lac promoter can “leak,” meaning that a low level of expression exists even without... -
Deep Dive: qPCR
TypeBlog Post...appropriate choice (Panina, 2018). Look for studies specifically aimed at determining the best options for...Zhang M, Hu G, Fang F. Fudenine, a C-terminal truncated rat homologue of mouse prominin, is blood glucose-... amounts of reagents in your chemistry, unless running a multiplex reaction or on a rapid cycle machine... quick way to validate your primer probe is by running your qPCR product on an agarose gel, then cutting...real-time RT-PCR. J. Immunol. Methods 283:291–306.Panina, Y., Germond, A., Masui, S. et al. Validation of...Digital Droplet PCR for AAV QuantificationPlasmid Cloning by PCR... -
Delivery Methods for Generating iPSCs
TypeBlog Post...compared to other delivery methods. Retrovirus has a cloning capacity of ~8 kb, so either a single polycistronic...dividing and nondividing cells and have a larger cloning capacity (8-10 kb) than other retroviral vectors...between a vector and chromosomal DNA. PB has a cloning capacity of ~9-14 kb. This system is transfected...transfected into cells as a donor plasmid containing the transposon, along with a helper plasmid that expresses...which is removed by culturing cells at 38℃, but screening cells for the presence of transgenes is still ...help mediate their transduction, such as the polyarginine peptide described in Zhou et al and Kim et ...large nucleoli, and little cytoplasm. Positive staining for alkaline phosphatase, expression of embryonic... -
Hot Biosensors 2022: Year-End Roundup
TypeBlog Post...dynamics in live mice under a variety of behaviors, opening up new research directions that were inaccessible...concentrations in cells than existing sensors. Using genome mining and structure-guided engineering of potassium binding.... Figure 5: Structure-guided design and genome mining both led to new sensors with lower binding affinities...sensor plasmids here! Torres Cabán CC, et al. Tuning the Sensitivity of Genetically Encoded Fluorescent... Indicators through Structure-Guided and Genome Mining Strategies. ACS Sens. 2022 May 27;7(5):1336-1346...cellular concentrations of lactate. The microfluidic screening platform developed here is also a promising approach...Figure 6: A) Schematic of BeadScan platform for screening candidate biosensor variants for performance under... -
What's New in CRISPR - Spring 2019
TypeBlog Post...plasmids String assembly gRNA cloning (STAgR) String assembly gRNA cloning (STAgR), developed by Stefan...Read the paper in Genome Biology Find plasmids containing these base editors Spatially and temporally...gRNA multiplexing system that generates vectors containing high numbers of gRNAs in one overnight reaction... Read the paper in PLoS One Find the STAgR cloning plasmids Mobile-CRISPRi allows CRISPRi experiments...Cas12a inhibitors Using a bioinformatics and a screening approach, the Jennifer Doudna lab identified three... -
CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More
TypeBlog Post...contain BsaI sites for easy cloning of a gRNA of interest. Cytidine and adenine base editing in Streptomyces...breaks are lethal because the non-homologous end-joining (NHEJ) repair pathway is not very robust. In many...applications for which they’ve been used. The beginnings of bacterial CRISPR engineering One common way...template, using Gibson Assembly or restriction enzyme cloning. Streptomyces bacteria are more recombinogenic ...directly modify a given gene. CRISPR-tranposons By combining CRISPR editing and transposons, the Sternberg ...inducer, in this case anhydro-tetracycline (aTc). By cloning in two tandem copies of the gRNA cassette, one ...Sun lab demonstrates the utility of cytidine and adenine base editors that enable C-to-T and A-to-G base... -
Viral Vectors 101: Integrase-Deficient Lentiviral Vectors
TypeBlog Post...might be considered a double-edged sword with unintended consequences. In this blog, we’ll take a deep...the name suggests, these lentiviruses lack a functioning integrase, the enzyme responsible for inserting...homologous recombination or non-homologous end joining (see Figure 2). Figure 2: Schematic representation...interruptions. Insertional mutagenesis is especially concerning in clinical settings, as evidenced by retroviral... -
Tips for Getting a Faculty Position
TypeBlog Post... Chalk Talk These items should be part of the beginning of your chalk talk. These should be plainly stated... implicated in several human diseases. Proper patterning of cells and tissues is fundamental to development...you understand what it takes to be successful in running a lab, getting grants, and writing papers? This... and earn some cash for my expensive habits of running shoes and single malt whiskies, but I remember ... -
CRISPR 101: Homology Directed Repair
TypeBlog Post... primarily by two pathways: Non-Homologous End-Joining (NHEJ) and Homology Directed Repair (HDR). NHEJ...your site of interest, and a donor DNA molecule containing your desired edit are all you need to generate...generate a knock-in. General considerations for designing a repair template to create mutations HDR donors...Featured Topic Page Learn about Non-Homologous End Joining Learn about Base Editing Read 3 Tips to Improve... -
Harnessing Bacterial Toxins for Allelic Exchange
TypeBlog Post...backbone. This step requires “counter-selection,” meaning that the selection is for the loss of a specific...protein. This allows conjugation between a pir-containing donor and the recipient of interest. Because ...transcriptional readthrough into the MCS to facilitate the cloning of toxic products. To allow manipulation of naturally...But if they are blue or magenta, they are real! Planning an allelic exchange experiment? Find the pTOX ... -
Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection
TypeBlog Post...protein. These were either (i) coded by the multiple cloning site and therefore consist of random amino acid... often use restriction enzymes in the multiple cloning site for generating fusions. For the POI-fluorescent...PVAT, when we use the AgeI site in the multiple cloning site) is sufficient. For the fluorescent protein-POI...immunofluorescence or properties of the protein. Determining other aspects of functionality (catalytic activity... -
Treating Muscular Dystrophy with CRISPR Gene Editing
TypeBlog Post...essential for proper muscle function. Without functioning dystrophin, an individual experiences progressive...skipping proceeds through the non-homologous end joining (NHEJ) pathway. It’s estimated that very little...Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F. In vivo genome editing using...Used to Edit Genes Read How Non-Homologous End Joining is Used to Delete Genes Additional Resources on... -
An Addgene Summer Internship
TypeBlog Post...expand my knowledge more than ever before. Before joining Addgene, I had spent most of my junior year stressed...well as complete the lab safety and cybersecurity training before stepping foot into the lab itself. I appreciated... as equals, because as soon as I had passed my training, I dove straight into my capstone project. A capstone...wrong. My capstone project was based on plasmid cloning and I, the researcher, got to choose what gene ... winner. We used the next 4-5 weeks to work on cloning the Syntaxin3 gene in a pLenti vector backbone....assembly but decided to go ahead anyway. We were running on a time crunch and knew that if we were to restart... -
Viral Vectors 101: Systemic Capsids
TypeBlog Post...Stauffer, W. R., Fish, K. N., & Pfenning, A. R. (2022). Machine learning sequence prioritization for cell...cells in C57BL/6J mice, but not in BALB/cJ mice, meaning that systemic capsids that rely on LY6A interactions...2017). AAV-PHP.eB has frequently been used for screening candidate regulatory elements such as enhancers...capsid of choice for future promoter/enhancer screening. However, AAV-PHP.AX packages viral genomes at...Gradinaru, V. (2017). Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous... -
Trading Spaces - Visiting Scientist Edition
TypeBlog Post...If you’re interested in learning a new skill, getting access to equipment you don’t have, or maybe both...researcher remains employed by their home institute, meaning the host university does not hire or pay them (...current institute. A common reason is hands-on training in a technique that only a few labs have expertise...this expertise will provide you with guidance and training so that you yourself can become an expert and ... Identify reagents you will need, any required training, and a priority list of experiments you hope to... Visiting scientist positions take commitment, planning, and funding – they may not be right or necessary...