... up a large and diverse group of professionals running the marketing, commercialization, collaborative...client might need help with the intricate task of designing a strategic vision for a portfolio of candidate...edge; a consultant might outline the impacts of combining targeted molecular therapies for cancer with chemotherapies...from previous consulting experiences, educational training, and conferences, and synthesizing this knowledge...externally to clients and experts. Hence, active honing of communication skills is a must to be successful...have greatly benefited from and appreciated the training I’ve received in concisely and accurately communicating...and that they can be ameliorated by thoughtful planning and engagement, leaving mental energy to enjoy...

...transitions are edits between the purine bases (adenine, guanine, and hypoxanthine) or between the pyrimidine...gene editing. Cytosine base editors (CBEs) and adenine base editors (ABEs) chemically modify target bases...included in the CGBE. They also developed a machine learning model capable of predicting editing outcomes of...sometimes processed by error-prone non-homologous end joining (NHEJ). To mitigate this, the team introduced the...of unwanted DNA edit. NHEJ Non-Homologous End Joining An error-prone cellular process to repair DSBs.... the first type of base editor invented. ABE Adenine Base Editor Converts A → G; the second type of ... screens, target-library analysis, and machine learning. Nature Biotechnology, 39(11), 1414–1425. https...

...that many researchers feel that they lack formal training in a variety of transferable skills. At Addgene...undergrad working with cancer cell lines. As I started planning out my own experiments in graduate school, I had... into three conceptual chunks: prioritization, planning, and procrastination. Time management through ...one of them. That’s why, as I’ll discuss in the planning section below, it’s important to dedicate specific...splitting my cells first. Time management through planning I probably wouldn’t have fallen into my cell splitting...projects to operate this way now. Talking about “planning” is easy, but actually doing it in a productive... way can be quite difficult. In my experience, planning has 3 components: A mechanism for blocking off...

... methyl group to the adenine in AACNNNNNNGTGC or GCACNNNNNNGTT Impact on cloning and digests Although...naturally occur in bacteria for purposes other than cloning or confirming plasmids. It turns out that restriction...one or a few restriction modification systems containing methyltransferases that add methyl groups to ...DNA: Dam methylase adds a methyl group to the adenine of GATC stretches of DNA Dcm methylase adds a methyl...coli strain and will therefore have methylated adenines in any GATC sequences found in the plasmid. When...enzyme will cut? Whether performing a digest for cloning purposes or for diagnostics, we suggest double ...can ensure this site remains unblocked by first cloning your DNA into a dam–/dcm– strain of E. coli such...

...Read about Other Plasmid Cloning Techniques like SLIC and Golden Gate Cloning Learn How to Verify Your...same efficiency as Gibson Assembly and GeneArt cloning - but at a much lower cost. Read on to find out...an appropriate vector. Gibson Assembly, GeneArt Cloning, and SLIC can all be used to assemble multiple ...described above, you design two primers, each containing the desired mutations, to amplify both strands...strands of a plasmid template. In addition to containing the mutations, the primers also have some homology... head-to-head with Gibson assembly and GeneArt cloning, the results are favorable. With 17-bp homology... observed efficiency of mutagenesis (mutation-containing colonies/total colonies) is about 84% for each...

...Positive Repressible: An activator protein is bound, turning transcription ON. When a repressor binds the activator... herpes simplex virus. tTA binds to promoters containing TetO elements (often linked in groups of seven...change that prevents binding to the promoter and turning transcription OFF. Despite their bacterial origins...systems function well in mammalian cells, and TRE-containing promoters can be used in the repressible manner...activation domain VP16. cTA binds to promoters containing putative operator sequences (CuO) (linked in ...change that prevents binding to the promoter and turning transcription OFF. The cumate switch system functions...broad QF expression-induced lethality in flies. Riabinina et al. also developed chimeric transactivators...

...system), in 2009 (at the beginning of the financial recession) I knew obtaining work in scientific research...Working in industry: Microbiology On returning from Ghana at the beginning of 2010, I explored options for ...physically carrying out experiments like PCR and running gels, but in research you also experience a lot...positive experience. I enjoyed helping people and learning to explain concepts in ways that people from another...looking for other career options after 6 months. Returning to university to study a Masters of Science I ... saved money from my role in Industry and took evening work to help cover the costs. My MSc courses were...Industry was really valuable; I had received all the training and experience relating to good manufacturing ...

...gets “Lexicon Training.” Since the lexicon is a “living” document, I update the training frequently. In...the confusion in our meetings. A lot of our LIMS planning involved “plates,” and our scientists and lab ...misinterpretation, that misinterpretation was definitely happening. Our solution for this problem was the creation...simplified the first weeks or months when I was learning the terminology of a new field. Annotate: When...we define new and potentially ambiguous terms. Defining words and applying annotation can be particularly... meeting is using the same words with the same meanings consistently, meetings will be shorter and more...Happier colleagues. Feeling like people aren’t listening to you or understanding you can be incredibly ...

...starts by determining what features you expect to differ between plasmids, and selecting or designing a primer...antibiotic resistance, restriction sites used during cloning, and most importantly, the predicted full plasmid...quality, as it will catch any errors made during the cloning, sample handling, or sequence assembly steps. All...on the number of features we need to verify. Determining which features to verify will depend on the particular...its predecessor. Important features may include cloning junctions (where most sequence assembly errors ...mutations, in-frame protein fusions, and multiple cloning sites. In the simplest case we have a standard ...you design your custom primers. Remember, when designing a sequencing primer, you need some space between...

...Arabidopsis in Education Learning Synthetic Biology with BioBuilder 3D Models for learning about CRISPR Resources...students how to perform these experiments in the beginning. In most classrooms, the laboratory experience...experiments. Notably, this plasmid harbors a multi-cloning site flanked by T7 promoters to allow for bidirectional...riboprobes) with modified nucleotides (for example, Digoxigenin-labelled Uracil) that will bind to complementary...complementary DNA (cDNA), and then use PCR and molecular cloning to insert the cDNA into the desired plasmid backbone...siRNA and are directed by complementarity to the remaining single stranded siRNA to bind natively expressed...-amputation planarians after knockdown of beta-catenin. Other gene targets do not require amputation to...

...just for lab specimens — there is evidence that lightning can also promote gene transfer in soil bacteria...resistance-mediated selection is indispensable for cloning work, antibiotics and plasmid-encoded resistance... in the lab but also causes outbreaks of food poisoning, the difference is virulence plasmids! This category...natural inducible expression systems that allow tuning of metabolic pathways. Fertility (F) plasmids: ...2024), and others have proposed developing new cloning techniques based on these vectors (Shareck et al...have more than one function: for example, Gateway cloning relies on genes found on an E. coli fertility plasmid...this F plasmid is critical for successful Gateway cloning. In addition, scientists have engineered entirely...

...something, even if that thing is opening a pizza joint. Silly ideas (mining local salt deposits to sell at...with a similar conundrum – she had the skills, training, and motivation, but there very few jobs available... journal clubs outside of work. Our scientific training was put to use as we explored ideas, planned out...” We kept our day jobs and worked weekends and evenings on the project, but still continued to generate...and another $30,000 in mentorship and business training. The money is a drop in the bucket against the... the technology we will need, and the business training to make the most of it. It has also allowed us...and beautiful, to engage with the public in a meaningful way about science, and to have a job that we ...

...ones? Here, I’ll cover some screening methods to identify them. Visual screening Regardless of the RNAi-trigger...organisms. When supplemented with adenine analogs such as 2-fluoroadenine (2-FA), organisms with functional...for viability (Morgens et al., 2016). Only upon combining the data generated from both methods did a more...easy creation of long hpRNA with Gateway-based cloning and straightforward positive selection of plants...assembly of intron-spliced hpRNAs through Golden Gate cloning. Artificial micro RNA and short hairpin RNA The... researcher, the traditional hpRNA and visual screening method was limited in two substantial ways: variable...morphological parameters, can be easily scored. However, determining the transcript or protein abundance of your specific...

...pHelper plasmid containing Ad E2A, E4, and VA RNA genes; and the pRepCap plasmid containing replication factors...outcome and planning.  Both ddPCR- and qPCR-based titering assays measure physical titers, meaning the concentration...those will quantify both genome-containing (full) and non-genome-containing (empty) capsids, thus leading...TCID50 measurement reports an infectious titer, meaning the concentration of viral particles that can transduce...concentration of viral particles containing a viral genome. While ddPCR is superior in precision and variability...accessibility of instruments in standard labs. When determining titers by qPCR, we recommend using primers targeting...

... be as pure as for other uses.   Cloning grade DNA Molecular cloning includes a set of techniques that...extraction. Figure 2: Use cloning grade DNA for direct cloning of your gene of interest into a ...in 3 different ways: Transformation grade DNA Cloning grade DNA Transfection grade DNA Figure ...can have a plasmid or viral origin. The type of cloning approach you choose ultimately will influence the...DNA (ng range). To allow efficient and flawless cloning it is important that the preparation is pure from...the Addgene blog Browse blog posts on plasmid cloning Read our Plasmids 101 blog posts Learn about the...

...professionals is learning to succeed in grants through trial and error, through winning and losing funding...may consider earning a degree in Grant Writing, Management and Evaluation and/or earning the GPC credential...grant writer’, but make no mistake, researching, winning, and effectively managing grants involves much ...consulting firm, a Twitter Chat, and then our grant training and mentoring. How do I become a grant professional...to help you enter and to have a successful and meaningful career in the grants profession: 1. Build on ...There are many grant consultants who offer grant training, mentoring and coaching services, including Smartegrants...entering the Grants Profession. Whether you’re transitioning from a previous career or you’re starting your...

...Scientists enjoy learning new things. Building new relationships is all about learning new things from ...Training as a scientist in the academic system has many pluses. I delighted in my graduate school years...nature of academia often means that scientists-in-training rarely get the opportunity to learn about the ...science. I find it ironic that we do all of our training as scientists (5-12 years worth!) with academic... one of the best things you can do during your training is meet interesting people doing interesting things...Informational Interview it is understood to be a nonthreatening request to discuss experience, interests and...really, always). Informational Interviews are for learning and for expanding your web of connections. A mistake...

... explore beyond the expected. By widening your search and maintaining an openness to unexpected possibilities...simple steps to welcome these unforeseen moments. Widening your search The first step towards inviting serendipity...ultimately enhancing your project's potential. Widening your search creates fertile ground for serendipitous...could spark unforeseen connections and ideas. Maintaining openness and looseness of spirit The second crucial...element in harnessing the power of serendipity is maintaining an openness and looseness of spirit. Amidst the...of inspiration. Additionally, journaling in the morning has proven invaluable for me, allowing me to pinpoint...think beyond the confines of a predefined path, opening doors to novel ideas and approaches. Moreover, ...

... multiple cloning site to insert YGOI, a bacterial selection cassette for growth and cloning in bacteria...conditions. The first thing you'll need to decide when running your experiment, is which vector will best suit...bells and whistles to your construct. If you are planning to track your protein in the cell or purify it...expression vectors are inducible (e.g. pBAD LIC cloning vector (8A)) and have an epitope tag (6xHis or ...expressed protein with a specific resin (e.g. 6xHis containing pDEST-HisMBP). After purification the protein...pBV-Luc) and gene silencing (e.g. pLKO.1 - TRC cloning vector). Check out our complete guide to empty ... Plasmids 101 blog series Addgene's Molecular Cloning Guide & Other Plasmid Protocols List of common ...

...option to indicate separate promoters, tags, or cloning information for each insert.  Encodes gRNA/shRNA...sequence. Empty backbone A plasmid designed for cloning a gene/insert into it. Empty backbones often contain...proteins that are appended to new inserts through cloning. *Note — If you will be depositing multiple plasmids... information about the gene/insert(s). Step 3: Cloning Information Please enter the name of the vector...vector backbone information. List any promoters, cloning details for how your gene was inserted into the...for each insert individually.   Figure 9: Add cloning information for the gene or insert. Step 4: Growth...possible, we propagate plasmids in the standard cloning strain DH5α. Please indicate if your plasmid cannot...