...studying membrane dynamics, for monitoring cargo trafficking, and for discovering organelle contact sites. This...signaling pathways to the cell cycle. Good tools for measuring intracellular pH are required in order to determine...endogenous gene activation. This system works by bringing together an anchor protein and activator protein...MCS/SacI site in pUCXKT, removing the stop codons during backbone digestion and providing the missing codons...utilizes the bulldozer-like quality of the ribosome during translation, which knocks off all mRNA binding ...NLS-MS2-RFP) the reporter mRNA, which contains the tethering sites for the fluorescent proteins and an inducible...both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins...

...presentations and a handout on protocol sharing, material sharing, computational reproducibility, and tools...wet lab protocols, reagent sharing, bioinformatic tools, data and code sharing, and data analysis and visualization...identify the right opportunities, he was able to bring together talented and motivated groups and individuals...s contributions by Tyler Ford included reagent sharing and were presented at the ASPB meeting by the collaborators... exclude different modules for this workshop. Empowering others to have the materials and resources to... everything that you would want to know before, during, and after organizing such a workshop. Finally,...Finally, the group put a lot of effort into ensuring that the content of the materials and the workshops themselves...

...genome engineering workhorse You rarely hear the word ‘CRISPR’ without shortly thereafter hearing ‘Cas9...Cas family of proteins has many useful genome engineering enzymes, each with their own unique features ... and quirks. Here, we will review the genome engineering abilities of the lesser-known Cas enzymes while...Cas9’. The Cas9 nuclease is a genome engineering powerhouse, and for good reason. It is guided by an RNA...strand break. Cas9 has been studied for genome engineering purposes far more than any other Cas family protein...making Cas9 an adaptable choice for many genome engineering endeavors. For more information on Cas9 variants...particularly convenient for targeting experiments requiring single strand DNA generation, such as homology...

... and Laura O'Regan Ordering jeans online from your favorite store is easy. Ordering genes on plasmids ...to other ordering questions here. In addition to answers to common questions about the ordering process...most frequently asked questions related to the ordering process: I placed an order through my university...will be able to enter your PO number as payment during checkout. Placing your order online has some benefits...Add all your items to your cart. Click checkout. During the payment step of checkout, you will have the... invoice to the billing address that you listed during checkout. This invoice will reference your Addgene...Addgene order number and PO number that you entered during checkout. At this time, your organization will ...

...approved 91,023 during the first 3 quarters of 2017. As an international researcher and aspiring entrepreneur... has been a growing shift away from employers sponsoring individuals for green cards because of the cost...have different roles, titles, and responsibilities during their careers - being aware of this can help take...regulations define a "specialty occupation" as requiring theoretical and practical application of a body...limited to biotechnology, chemistry, architecture, engineering, mathematics, physical sciences, social sciences...business specialties, theology, and the arts, and requiring the attainment of a bachelor's degree or its equivalent...work-authorization is strictly limited to employment by the sponsoring employer. On April 18, 2017, President Trump ...

...vector into the culture medium during adeno-associated virus manufacturing. Human Gene Therapy, 21(10), ... iodixanol) using a high-speed ultracentrifuge. During the spin, the virions will form a band due to their...check out the Addgene AAV Production protocol. Titering Once you’ve produced, harvested, and purified ...titer of your viral vector solution. A variety of titering methods have been developed to determine the titers...TCID50) (Zen et al., 2004).  To choose the optimal titering assay, you’ll need to understand how different...outcome and planning.  Both ddPCR- and qPCR-based titering assays measure physical titers, meaning the concentration...has to be digested with DNase before undergoing titering, to remove residual plasmid DNA carried over from...

...worry about traveling during an already stressful time, and I was more relaxed during both sets of interviews...yourself to a few members of your desired department during the application process. Many institutions recommend...in case one isn’t able to respond to your email. During my PhD application cycle, I reached out to around... and make sure you cover everything you need to during the interview. (Virtual) Campus Visits After receiving... financial and logistical obstacles could make touring potential programs and universities stressful or...Through the virtual systems created and promoted during COVID-19, online meetings with current students...worked with students to maintain these time goals during COVID-19? How are students funded? Are there any...

...genome engineering across multiple species Recent discoveries in the fields of genome engineering and synthetic...multiplex automated genome engineering (MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate...gene of interest. Although the parts involved in altering gene expression are relatively well known, particularly...labs who wish to harness this powerful genome engineering tool. Bassett, et al. J Genet Genomics. 2015...loci (multiplexing). Currently, the only genome engineering method in bacteria that enables rapid, automated...combinatorial genomic libraries. MAGE has allowed genome-engineering endeavours of unparalleled complexity in Escherichia...mutant enables transient suppression of DNA repair during oligonucleotide integration, allowing MAGE in otherwise...

... with limited time for one-on-one mentoring. His solution? Bringing his research into the classroom. I...textbooks” (Brian, 1996). But is it possible to bring the kinds of transformative educational experiences...dehydrogenase (MDH), a citric acid cycle enzyme he studied during graduate school. As a stable, easily purified protein...interest to other researchers in their field. Indeed, sharing results with the broader scientific community represents...all shaped his teaching style, with the former offering excellent training in improvisation and the latter...Jessica Bell, Lisa Gentile, Ellis Bell, and Amy Springer), this group now connects MDH-focused CUREs at...available technical and pedagogical resources, offers mentoring for instructors, and provides opportunities for...

...community. Sharing is caring We’re thrilled that our depositors reap the benefits of sharing plasmids with...hooked on sharing because, as UNC’s Bryan Roth says, Addgene has “literally enabled sharing with 100's...plasmids deposited at Addgene enhance scientific sharing worldwide but a select few are particularly special...honor the creativity, passion, and dedication to sharing epitomized by the creators of these plasmids with...appreciate our depositor’s dedication to scientific sharing, we wanted to celebrate our depositing PI’s who...deposited in Addgene!” As you can imagine, answering plasmid requests can be a time-consuming chore...Flame Award winners- we’d like to thank you for sharing your plasmids at Addgene. You are the reason we...

...mix. During the incubation the immobilized antibody binds to the target protein, thereby tethering it to...approaches allow for optimized strategies for differing amounts of target proteins or antibody bonding...immunoprecipitation and simply refers to the process of anchoring an antibody, often called the capture antibody...eliminating the risk of the primary antibody interfering with downstream assays. This is particularly ...for an IP, samples are typically lysed in a non-denaturing buffer containing non-ionic detergents, such ...proteins, including those in the nucleus, more stringent buffers may be needed. Since many factors, including...Frequently, the antibodies used for a standard, denaturing Western blot will not be suitable for an IP, ...

...team building and bring back some degrees of belonging that was probably lost during these weird times...Discussing data with colleagues can be more difficult during the pandemic era, with fewer opportunities for ...   Implement new tools for creative interaction During Zoom meetings, different tools can be used to promote...The annotate function can also make the screen-sharing experience more interactive. The button is on the...collaborative cloud tools such as Google Docs and Sheets during Zoom lab meetings to chip in ideas and take minutes.... This document can be shared and serve as a springboard for future discussion. Collaboration and creativity...great functionality that is widely used and could bring back a taste of that casual kitchen chatter, ultimately...

...immune system to fight progressing cancers. It was inspiring to participate in the 2017 Cell Symposium on Cancer... are coming together to tackle the challenge of curing cancer in cross-disciplinary projects and collaborations...reflect how cancer/immunology researchers are preparing and carrying out this multi-pronged attack. Inflammation...our recent post “Cancer and the Immune System: Deciphering the Relationship” by our guest blogger Subhadra...CRISPR/Cas9 methodology with T cell receptor engineering (Ref 5)  as described by Blue Flame Addgene depositor...Professor Michel Sadelain from the Memorial Sloan Kettering Cancer Center. In contrast to using randomly integrating..., CRISPR/Cas9 genome editing allows targeted engineering of the TCR locus. He also highlighted the UCSF...

...to all of our generous, sharing depositors. Over 3,000 laboratories are sharing plasmids (with 16 labs ...labs currently sharing viral preps) through the Addgene repository. Without these scientists and the support...to have such an impact on scientific materials sharing. I would also like to recognize a few scientists...Addgene has been successful in helping thousands of sharing scientists from laboratories all over the world...Scientist Team has been developing expertise in answering questions on virus and helping provide educational...reach scientists around the world. Our Software Engineering Team (and, added in 2018, our Product Team) continues...Maintaining our excellent service and accuracy involves caring and careful Addgenies, but also a lot of good tracking...

...the work by using the principles of recombineering. In recombineering, bacteria express phage recombination...REPLACR-mutagenesis, harnesses the power of bacterial recombineering to create insertions, deletions, and substitutions...how to replace your SDM method with REPLACR (Recombineering of Ends of Linearized Plasmids After PCR). ...modification by homologous recombination. In most recombineering strategies, only 3-4 phage proteins are needed...directly transformed into bacteria expressing recombineering proteins Red/ET from a temperature-sensitive...bacteria are subsequently grown at 37 °C, the recombineering plasmid is not maintained. Recombination efficiency... the need for selection usually required in recombineering. REPLACR-mutagenesis is also well suited for...

...their biological materials during funding transitions. Enabling resource sharing: Our operating model allows...agility as a nonprofit resource can prove to be vital during this period of constrained funding. Addgene is ...lost amid lab closures or project terminations, ensuring that scientific progress made through years of...structures and indirect cost caps more efficiently. Offering free educational resources: Our eBooks, videos...productivity and efficiency with fewer resources. Fostering new partnerships: We're seeking and expanding ...strengthening our commitment to collaboration and resource sharing, we can navigate this difficult period together...what support would be most valuable from Addgene during this time. Your input will directly shape our response...

...into their scoring functions. Many tools have also opted to mix-and-match different scoring algorithms... by comparing their past results to the designs suggested by each tool. For those who are entering sgRNA...facilitate most experimental applications in CRISPR engineering. When compared to that collective utility, it... and approximate methods. The details of their scoring function can be found in their online tool’s documentation...is by far the most common with most tools also requiring that the user specify an organism. Output diversity... genome”, “large genome”, and “organism biased scoring”. Trends: Many tools can handle genomes of any ...solution but are often packaged as offline tools requiring expertise in one programming language or another...

...consideration when titering a lentivirus is the cell line that is used for titering. Typically researchers... my (lentiviral) generation The first step in delivering genes via lentivirus is deciding what packaging... competence can arise from recombination events during viral production and poses a biosafety risk to ... plasmids in order to obtain the best titers. Requiring an additional plasmid for transfection can make... are required to generate RCL. To overcome the titering hurdles of third generation packaging systems,...will choose a relatively permissive cell line for titering in order to obtain the best titer possible. While... One should also consider the specifics of the titering transduction experiment. Media choice, reaction...

... and morphology, rendering their continued use inadvisable for experiments requiring reproducible results...visible and you should always ensure nothing is covering it. Turning on the UV light 3-4 times a week for...remove all jewellery as well as that watch you’re wearing before you start. Also, avoid unnecessary talking...hood. Planning your experiments thoroughly before entering the hood is another great way to prevent contamination...you’ll know exactly what materials you’ll need to bring into the hood and can wipe them all down before ...air flow around your work area. Make sure to only bring required materials into the hood. This will give...sure cells don't stay out too long While you’re preparing things, make sure your cells don’t stay out too...

...accomplish is to make the engineering of biology much easier. So you could imagine engineering the entirety of ...Stanford develops engineered DNA systems capable of storing data and computing inside living cells. He's also...an organization working to advance biological engineering openly so as to benefit all people and the planet... what we think of as an operating system for engineering an entire cell. Typically when people use plasmids...cell. Until we get a handle on precise, forward engineering of the central dogma at the genome scale, it'...collection of genetic parts for precisely forward-engineering the central dogma across many genes simultaneously...been 40 years since the invention of genetic engineering. What's incredible is the bioeconomy is already...

... generated will enter the culture medium during subculturing processes or from the air if the cell line...that are generated (while talking or pipetting) during working in the laminar airflow are also among the...source of the problem is the media, used for subculturing, which is available in both liquid and powdered... before sampling (Uphoff and Drexler, 2011). Culturing on agar plates/broth is considered the ‘gold standard...separate lab should be made available for cell culturing and maintenance of the cell lines. Providing such...coats. Have separate footwear for the lab to avoid bringing in environmental contaminants.  Figure...Addgenie works in the tissue culture hood while wearing PPE. Photo by Kate Harten DeMaio. Cell lines...

... cell or neuron type may not work well for a neighboring cell type, even within the same brain region ...What AAV vector is best for my specific system?  Sharing data and community concerns AAV vectors are complex...particular importance to these efforts is the open sharing of negative data.  Negative AAV data is highly ...why they don’t share, many express concern about sharing negative data they cannot explain, perhaps out ...therefore actively work to remove the stigma around sharing negative data and instead focus on getting this...if many scientists choose to help each other by sharing their data. So take advantage of it and get your...Lukner, H., Kogermann, K., & Sepp, T. (2021). Data sharing practices and data availability upon request differ...

... January 2025.  What will the ordering process look like?  The ordering process is split into two steps...   Figure 1: The process of requesting and ordering a Packaged on Request viral vector prep.  ... research and expanded choice when it comes to ordering high-quality materials from the Addgene repository...Retrograde (AAVrg) capsids.  Please be aware that this ordering process will look slightly different from our ...steps, requesting and ordering. This split allows us to provide researchers with increased choice and flexibility...flexibility; while ensuring we can maintain high standards for both our depositors and requestors.  Requesting.... There is no charge for the request process.  Ordering If your request is approved, you will receive ...

...Wisconsin lab for a summer during my PhD and is responsible for my enduring love of cheese curds. We’...long line for bag check was definitely worth it! Hearing about his passion for global health and the various...one of the highlights of the conference. It was inspiring to see how microbiology research is improving ...researchers who weren’t aware of Addgene’s plasmid sharing services, as well as #happydepositors who stopped...leverage their expertise as we advance our mission of sharing science. I got to check out many of the posters...about our mission of open science and resource sharing and was also a great place to catch up with former...distribution service! Did you see a must-have plasmid during ASM? Let us know what’s missing in the collection...

... she's either day-dreaming about exploring sci-fi worlds or exploring nature in the real world. Additional...ICE on Synthego’s blog. CRISPR-based genome engineering has revolutionized the gene editing field by ...edits performed over multiple experiments and comparing the robustness, accuracy, and speed with existing... of the sample name. If there was a minor error during processing, the window shows a yellow checked circle...with non-wild type sequence) as determined by comparing the edited trace to the control trace. In the ...Value - When the ICE linear regression is computed during generation of the ICE Score, the Pearson correlation...sizes in the entire edited population of genomes. Hovering over each bar of the indel plot shows the size...

... Figure 1: Addgene’s Agile Approach Sharing is caring There are many moving parts to our software...overall soundness of the software design. Each of us brings slightly different cognitive biases as a result...defined chunks, called stories, and work on stories during short periods of time called sprints. The team ...different technical approaches that may come up during discussion.  One example of agility at work is ...we recently reduced the number of steps involved during checkout. Demo details for this would be something...diversifying our own knowledge of the software, sharing knowledge allows for life to happen without impacting...developing new features and fixing bugs: we take time during each sprint to do some maintenance work on our ...

...without the undesirable side products generated during chemical synthesis of these molecules. To make ...recombinant protein purification and metabolic engineering. If instead you want to build genetic circuits...biology MoClo applications, but there are more! Engineering disease resistance through gene stacking, increasing...kit has a focus on transformation and genome engineering. This is a specialized system with parts including...Multiplex MoClo Toolkit for Extensive and Flexible Engineering of Saccharomyces Cerevisiae.” ACS Synthetic Biology...Mitochondria.” BMC Biotechnology, vol. 17, no. 1, Springer Science and Business Media LLC, 13 Nov. 2017. ...Vector Toolkit and Parts Library for Advanced Engineering of Plant Genomes.” The Plant Genome, vol. 16,...

...combines CRISPR and recombineering-based MAGE (Multiplex Automated Genome Engineering) technology. pMA7CR... a backbone-targeting gRNA cassette for plasmid curing through "self-destruction" after induction with...CRMAGE is much more efficient than traditional recombineering, with 96-99% efficiency for point mutations...for a single round of editing, and a subsequent curing protocol that requires only 2-3 hours incubation...References 1. Maddalo, Danilo, et al. “In vivo engineering of oncogenic chromosomal rearrangements with ..., et al. “Multiplex CRISPR/Cas9-based genome engineering from a single lentiviral vector.” Nucleic Acids.... Sakuma, Tetsushi, et al. “Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector...

...repair and genome engineering CRISPR-Cas9 has revolutionized the genome engineering world and made targeted...Originally written by Marcy Patrick and Mary Gearing on Mar 12, 2015; updated by Christina Mork, Jul ... fill in the resulting ssDNA. A single HJ forms during this process and resolves by cleavage of the crossed...immediately applicable in DSB-induced genome engineering, it has biological importance for the repair ...intact. In other words, Cas9 will keep cutting and repairing your target DNA until a mutation is introduced...Homologous Recombination. Jasin M and Rothstein R. Cold Spring Harb Perspect Biol (2013). PubMed. DNA double-strand... (2018). Pubmed. CRISPR References: Genome engineering using the CRISPR-Cas9 system. Ran FA, et al. ...

...other to be physically present at certain times during the day? We asked three lab heads how they go about... If you are looking to join a lab, you may be wondering “What is this PI looking for in a new researcher...of CVs collected from throughout the year since hiring opportunities arise at various times. If the timing...will provide a solid foundation on which to thrive during time spent as a graduate student or postdoc. Although...chosen research field is important, the end goal during this time is to be trained to think and execute.... Or perhaps, the lab has a reputation for not entering collaborations. Alternatively, perhaps the lab...to ask lab heads, lab members and lab neighbors during the interview process. What if I want a non-academic...

...some of these questions in the sections below. Mentoring relationships can help identify your priorities...Alumni meetings can be the most informative way of gathering data. Former lab members can give open, revealing... Know who's boss You sign on to a lab for the mentoring only to find out that the lab head is an absentee...You are not allowed to speak to lab members alone during the interview period. People seem to be hiding ...about discussing data. There’s hesitation before answering questions about lab culture. Support for diverse...ensure that you are expanding your options and preparing for both academic and non-academic career paths...evidence of industry collaboration? If you are considering a career at a small, teaching college you might...

...methods for physical shearing exist as well, such as sonication and needle shearing. Essentially, any method... with the lab during its recent move to the University of Georgia). She enjoys pondering how phages get...between 1 and 3 kb results in optimal likelihood of capturing a whole RBP gene without causing issues associated...following colony PCR of 10-20 colonies using primers bordering the multiple cloning site of the vector. In order...saturate each with lysis solution (~3 ml each), pouring off any excess. When your protein induction is .... Leaving the membrane overnight in the freezer during one of these steps is also a convenient stopping...through the day, spending time with her horse, and wondering what her cat is thinking. Find her on Twitter ...

...for many metabolic engineering and synthetic biology goals. In metabolic engineering, the expression of...interfere with the precision needed in metabolic engineering or other synthetic biology circuits. What is ... a look at how the stabilized promoters perform during these types of fluctuations. When cells are dividing... can also cause changes in plasmid copy number. During the shift from exponential to stationary phase,...promoters were able to better modulate gene expression during shifts in growth rate as compared to a constitutive...To simulate the increase in plasmid copy number during the transition to stationary phase, Segall-Shapiro...they could alter copy number of the plasmid by altering IPTG concentrations. Higher IPTG concentration...

...leads to incomplete reprogramming. Additionally, lingering expression or re-expression of viral transgenes...temperature sensitive Sendai virus which is removed by culturing cells at 38℃, but screening cells for the presence...considered the safest since there’s minimal risk of lingering expression of reprogramming factors. Non-replicating...the plasmid in mammalian cells and assists in tethering the vector to the cell’s chromosomes. With the... high marks for safety.   Protein Delivery: Delivering reprogramming factors as protein is another way... week for 6 weeks, and still required further culturing to generate iPSC colonies. Reprogramming efficiencies...three germ layers (ectoderm, mesoderm, endoderm). Measuring pluripotency of mouse cells typically involves...

... organisms which I use to study gene regulation during developmental processes, CRISPR/Cas9 allows us ...laborious handling (screening 768 F1 animals). Recovering rare editing events in a sea of unedited animals..., which makes screening extremely rapid. This stringent selection allowed me to optimize a range of editing...al., 2018). They further built on this work, discovering that melting and rapidly cooling dsDNA repair...also found that the majority of knock-ins were occurring 24 hours post-injection, making screening for ...where the 5’ ends of the PAMs are facing outwards, ensuring the PAMs remain on the chromosome after DSB formation...Toolkit and SapTrap-SEC kit for C. elegans genome engineering have also facilitated CRISPR editing in C. elegans...

...the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 technology has revolutionized the...the fields of molecular biology and bioengineering, as it has facilitated the development of a simple ...relative nuclease efficiencies as well as to more stringent targeting rules that result in a decrease in available... as a means of modulating gene expression (6). Altering the length of gRNAs targeting different sites ...Santa Cruz, where her research is focused on engineering nanoparticles as drug delivery systems. She is...develop several popular CRISPR-based tools for altering DNA sequence or regulating gene expression.   ..., Raj, et al. "Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach....

...proteins in solution. The pH is then lowered using a renaturing solution, which causes the proteins and genomic...purification and DNA gel extraction with glass syringe filters Starting material: bacterial cultures or...binding DNA to silica columns, it’s bound to glass syringe filters. Normally DNA does not bind silica or glass... kit takes 1-1.5 hours to complete, while this syringe-based method can be completed in 20 to 30 minutes...culture you’re working with. Overall, the glass syringe filter method is not only cheaper, but also faster...gel slices Product: DNA fragments You might be wondering, “How is milk going to help me extract DNA from... 48 kb with high yields and without degradation during the extraction procedure. Additionally, purified...

...make genomic alterations, bringing about a revolution in genome engineering, with new techniques rapidly...and can be used for most of the current genome engineering applications of CRISPR including: generating ...delivery. You’ve decided to use RNPs, now what? Preparing the Cas9-gRNA ribonucleoprotein The first step... the Jacob Corn and Alex Schier laboratories. Delivering Cas9-gRNA ribonucleoproteins  An advantage of...applications. This technique utilizes Cas9 proteins harboring receptor ligands (C in the figure above), which...stable in human plasma than the standard spCas9 (Harrington et al. 2017). Lastly, as with any scientific ... Addgene.  Andrew's interests include genome engineering and microbiology.     References 1. Chen, Sean...

...accelerating scientific discovery. Over 150,000 plasmids empowering researchers worldwide. Countless breakthroughs... to make it easier to find what you need and to bring it in line with current Web Content Accessibility...contributions to key research papers are already inspiring higher standards in antibody use (Ayoubi et al...packaged on demand in five distinct serotypes, offering you unparalleled flexibility. But this is just...characterization data for over 150,000 plasmids, empowering bioinformaticians, data scientists, and developers...collaboration formed, and every breakthrough enabled brings us closer to making an impact on humanity's greatest...this incredible journey. Together, we're not just sharing research materials — we're accelerating the future...

... Members periodically break, go exploring outside the team, and bring information back. No team has all...contribution short and sweet. One key to this is ensuring that the group always conducts exceptionally well-managed...manager can be efficient without delegating and empowering the team to solve problems together (tactics ...important paths of information flow is the one that brings feedback back to the manager. Team members rarely...often A one-time event doesn't work to get people sharing.   Frequently ask direct questions for specific...at formal communications events or at informal gatherings, like at lunch. Many experts recommend using ...constructive feedback and suggestions. Consider offering rewards (gift card, afternoon off, etc.) Act on...

...transferable skills while working in the lab and by offering advice on areas where you might need some help...important in a non-research context, especially when considering small to mid-sized companies and organizations...time I’ve had a role in organizing collaborations. During my graduate studies at Karolinska Institutet I ...what was expected, and keeping track of speakers. During your graduate or postdoctoral work there are loads...you develop will lead! Read our blog post about fostering communication in remote collaborations! Stay in...Great progress! You are collaborating with your neighboring lab, you've met interesting people at conferences...twitter and maintain a connection by commenting and sharing. LinkedIn or ResearchGate allow you to send personal...

... your foot in the door When I was first considering preparing for and transitioning to a career in healthcare... Healthcare consulting is a fast-paced field, requiring people who are willing to quickly learn, and apply...governments on the most effective and efficient ways of delivering healthcare. Clients typically engage consultants...fields. I got my PhD a couple of years ago in Bioengineering, and one of the things that pleasantly surprised... suite of benefits a given therapeutic approach brings to patients, doctors, and insurance providers, ...aspect of research in biology or medicine you will bring an essential set of tools to a consulting team. ...an increasing recognition of the value graduates bring to these roles. Healthcare consulting is a demanding...

...Read here to get help if you are having trouble preparing your most excellent resume. 2) Follow job boards...reviewed regardless of personal connections to the hiring group.  Read here for more on timely submission... job description is usually a wish list and the hiring manager knows that no applicant is likely to meet...Your resume must be in the official “system” for hiring managers to take next steps and follow up especially... as you will not likely know the exact person monitoring submissions (usually a Human Resources representative... your resume pdf document. 6) Use your network! During the entire process, use whatever means you can ...can to get your resume into the hands of the hiring manager on paper or as an email send directly from a...

...C., Addgene’s intern coordinator shortly after hearing her talk about Addgene’s internship opportunity...summer interns presenting their capstone projects during the Summer Intern Poster Presentation.  As ...the Syntaxin3 gene in a pLenti vector backbone. During those five weeks, I worked with my mentors, Katherine...always came with surprises. Originally we thought during the first three weeks that we had made a mistake...not having the right concentration of a reagent during our HiFi assembly but decided to go ahead anyway...restart the procedure we would not have enough time. During the following two weeks, we were able to obtain...on my capstone project report and presentation, during the final week at Addgene. This report shared my...

... assays are not with denaturing agents like DTT or β-mercaptoethanol.  Denaturing proteins To denature...blot, your loading buffer will not contain any denaturing agents. However, you’ll still have to lyse your... post, we are assuming that you are running a denaturing blot.  There are a number of lysing methods and... aliquot your sample(s) before adding in your denaturing and/or loading buffers, as most Bradford assays... proteins in different patterns, due to their differing chemistries. You therefore cannot compare Bis ...and reprobed multiple times. It doesn’t degrade during long-term storage and has both high binding capacity...well for proteins of most sizes and are worth considering if your proteins of interest vary wildly in size...

...research purposes, like creating stable cell lines, delivering CRISPR/Cas9 to cells for genome editing, and ... host’s genome isn’t uncommon. Many naturally occurring viruses occasionally integrate their genomes into...the host genome via the viral proteins packaged during production (read our blog post on the retroviral...time, IDLVs are ideal. As the episomes are lost during cell division, the expression of the delivered ...production protocol.  One thing to keep in mind is that titering these IDLVs may require a slightly different protocol...regular lentiviral vectors. This is because most titering methods rely on isolating genomic DNA from transduced...DNA along with a reference cellular gene. When titering IDLVs, there is no integrated piece to measure...

...be taken seriously so we made it official by registering BosLab inc. as a nonprofit organization in the...are skilled with audiovisual media, software engineering, business consulting, mathematics, and more. ... to perform proof-of-concept experiments for discovering novel therapeutics. Dave is a chemical biologist...and in his free time, he is pursuing a plant engineering project at BosLab. Once he optimizes plant cell...knowledge with the community by teaching a DIY plant engineering workshop. Community projects at BosLab Not all...and Mrina Nikrad have started the Open Yeast Engineering Project (OYEP). Through this project, community...do similar things. The focus is to make yeast engineering accessible to everyone but these projects could...

...problem solving. I believe that is something lacking during degree courses; they never really prepare you for...failure and problem solving scientists experience during research. Working abroad in rural Africa Once this...another area which I was really passionate about during my degree. I loved parasitology. However, when ... held me back; I do not have a second language. During the recession there were many qualified scientists...Genetics to give me an edge over other graduates during the recession. As each advertised job received ...the training and experience relating to good manufacturing practice (GMP) and good lab practice (GLP) (...recognised the logo, but didn’t know much else. During the interviews, I asked if I could contribute more...

...open science nonprofit that would facilitate the sharing of plasmids between scientists. The service would...resource. I really appreciate the work you do to make sharing reagents and protocols so easy!” — Nina Jain, Harvard...seven before we had 1,000 depositing scientists. Ensuring that people knew how to access Addgene, and that...2012, our repository was rapidly able to support sharing of CRISPR constructs and information across the...lentiviral and AAV vectors, as well as developing and sharing supportive educational resources. By 2017, we had...NGS run.   To twenty years and beyond! Which brings us to 2024, Addgene’s 20th anniversary and the ...been made because scientists believe in openly sharing their plasmids. It’s been a privilege to find and...

...head when you hear the phrase “action potential?” Firing neurons are likely the first thing that comes to...reprogrammed regenerating tissues without using genome engineering. Planarian worms cut into three segments “remember...communication is briefly inhibited in the entire worm during regeneration, the worm creates two heads. Subsequent...resources from the bioelectricity field In addition to sharing tools to study bioelectricity, bioelectric researchers...defects, regeneration, medicine, and synthetic bioengineering. References Chernet, Brook T., et al. "Use...growth and form." Annual review of biomedical engineering 19 (2017): 353-387. PubMed PMID: 28633567. Mathews...bioelectricity for regenerative and synthetic bioengineering." Current opinion in biotechnology 52 (2018...