... optogenetic tools in a deep part of the brain, making it sensitive to light. When they illuminated this...patients? Over a quarter of a million people have some kind of neural implant already. If we can use light to...maybe we could treat patients suffering from Parkinson's, chronic pain, deafness, or other conditions.... emotions, to see how a monkey changes decision making, to cure a human patient. We can't do it all by...you have any special tips or tricks for someone looking to try these optogenetic tools out for the first...always tell people, 'Be driven by your science.' Looking for Optogenetics Plasmids? Plasmids for Optogenetics...

...gene edit. I wanted to inactivate the gene BRAF (a kinase implicated in several human cancers) in A549 cells...Vinculin is a loading control. 3. In terms of making the monoclonal line: I had never done it before...found here. Figure 3: A surprise for the author. Checking Cas9 expression Overview: By Western blotting ...difficult part of the assay was designing primers flanking my edits in genomic DNA and doing PCR on genomic...agarose to get sharper bands. Conclusion Overall, making a human cell line with a gene knockout was straightforward...who you are, you could quickly be on your way to making new scientific observations. References 1. Rago...C, Vogelstein B, Bunz F. Genetic knockouts and knockins in human somatic cells. Nature Protocols 2007;...

...like multi-color imaging (Figure 2). If you’re working with purified proteins in vitro, you can even use...tags makes updating your labels easy! Instead of taking the time to clone a new construct for every new-and-improved...time points.   Figure 3: Single-molecule tracking of actin filament assembly-regulating proteins...developed over the years; improving the labeling kinetics or the fluorophore’s microenvironment to enhance...A., Juillerat, A., Heinis, C., Corrêa, I. R., Kindermann, M., Beaufils, F., & Johnsson, K. (2008). An ...Reinstein, J., Hiblot, J., & Johnsson, K. (2021). Kinetic and Structural Characterization of the Self-Labeling...Selecting Fluorescent Dyes and Ligands GCE4All: Making Genetic Code Expansion Accessible Addgene's Genetic...

...something difficult, be excited about the prospect of working toward a solution. At the end of the meeting, make...person to add a bullet or two about what they are working on. Building this into the agenda ensures that ...items, and Accountability Agendas: Send around talking points for the meeting a day in advance. This gives...Accountability: At your next meeting, start by checking in on the action items from last meeting. Use ...project moving. Use time wisely Meetings are for talking, not reading: Send around relevant documents, like... Executive Director Joanne Kamens FInd Tips on Picking a Research Project Read Other Inside Addgene Posts...

...the day you defend to start thinking about your next step. Start thinking now. Start browsing job postings...in your resume. If the job posting says “French-speaking is an asset,” mention “speaks French.” Employers...groups? Perhaps you had some supervisory experience working with undergraduate students? On top of that, perhaps...universal. Some universities even have officers working specifically to assist graduate students in job...postings and salary expectations public, so start by looking at these university postings to give yourself a...

...you build your lab team and foster an effective working environment? Tom Ellis was offered a start-up package...ideas, and ways to improve the lab. “An effective working environment for research is totally dependent on...communication amongst the team so getting everyone talking to each other cannot be undervalued.” Connie Cepko...of slowly building up your lab and selectively picking the first couple of hires to build a strong, collaborative...resources for Early Career Scientists, including their "Making the Right Moves: A Practical Guide to Scientifıc... Dr. George Church (Harvard Medical School) for taking the time to speak with us about adding new scientists...

... these files to your working directory (the folder that you are currently working in) every time you use...notebooks just won't cut it for bioinformatics. All kinds of biologists are finding themselves using computational... task will keep you from repeating your work or making similar mistakes in the future. Stay organized...comment in C++ and Java Command line history: Tracking command line history by recording all commands...system that does is through trial and error, and by asking for tips from others. Comment below to add your...

...Blog Mentoring for Scientists Series. I have been thinking a lot about Mentoring for over 10 years. Many ...Cepko to learn about her mentoring style. While working on this project, I read the book “Every Other Thursday...Form your own peer mentoring group In my travels speaking to grad students and postdocs I have started to...example, a group of people who are all interested in making a transition. Schedule your first meeting and get...breakfast once a month. All participants must commit to making the meetings a scheduling priority (and last minute...career paths and get their advice. Attend a local networking event together and help each other practice meeting...

...heavily with other skills such as critical thinking, decision making, creativity, or data analysis. Conveniently...researchers do learn MANY transferable skills while working in the lab and by offering advice on areas where...you spend time in a lab learning techniques and taking on independent projects, you will inevitably develop... skills passively. But I have always found that taking a conscious or active approach can be hugely helpful...are, while also practicing systematic, logical thinking.  If you are already a lab veteran at this point...

... Research. On December 1st 2016, many postdocs working more than 40 hours per week could expect to see... salaried workers receive overtime payment for working more than 40 hours per week increasing from $23,660... postdocs were at institutions focused only on tracking hours, and 22% of postdocs were at institutions...going ahead with their planned raises, some are taking the opportunity of the lapsed mandate (or are being...University of Massachusetts Medical School. We are working with postdocs at other institutions to see if we...that many postdocs already made plans and budgets working around issues such as childcare using money from...

... be useful. Mainly because, well, antibodies are kind of hard to see. To solve this issue, researchers...post, where you can find some additional tips for working with these tools.          Enzymes An equally, ...properties allow for efficient signal amplification, making biotin conjugated antibodies good options for detecting...lead perfectly blissful, productive lives without thinking about these chemical reactions. After all, there... are ready to use off the shelf. So why are we talking about this? Even though there are so many commercially...antibodies’ performance after the reaction.  If you are looking to conjugate antibodies yourself, there are kits...

..., consider making you own DNA ladders with these plasmids from Penn State Need help picking the right ...cells due to their IgGκ signal peptide sequence, making them easy to harvest from cell culture media. Their...HA staining is weaker than reporter staining. Picking the right reporter for your RANBody While all three...staining GFP, RFP, H2A2B   Are you ready to start making your own RANbodies? Great! All of the RANbody plasmids...learn more about nanobodies Additional Resources Looking for primary monoclonal antibodies to use with the...

...Novartis Oncology.  Bhang did her PhD work at Johns Hopkins and came to Novartis for her post-doc in January...after treatment with EGFR inhibitor, erlotinib. Looking at barcode abundances among multiple replicate ...Pharmacology group at Novartis Oncology, Bhang has been working on using the ClonTracer library to monitor clonal...their own work. She says, “I spend a lot of time packing, shipping, and sharing protocols one by one,” and...is a “really great way of broadly sharing these kinds of reagents.” Here at Addgene, we’re excited to ... to use! Many thanks to Carrie and Novartis for taking the time to talk with us and for providing this...

...started working in the pharmaceuticals industry after earning my Master’s in 2016 and then working four ...prepare 1-2 unique questions for each interviewer by looking at their LinkedIn profiles. This can help spark...your 1-hour presentation to your audience Start by asking the hiring manager or recruiter what presentation...improving a process or procedure, solving a problem, or making a key decision. Even deciding to stop pursuing ...sharing your screen and won’t be able to see who’s asking a question. Maybe you ask for questions after completing...

...proteins had to be re-engineered for each gene, making wide-scale gene activation seem next to impossible...activation uses dCas9, a CRISPR protein variant lacking its endonuclease ability, to bind to genes without...there are multiple options to choose from when looking to activate genes across diverse cell lines.   ...activation compared to other CRISPR activators, making it a popular method for gene activation experiments...design (SunTag). This streamlines its delivery, making it a common choice for CRISPR activation. Newer... Larson MH, Gilbert LA, Doudna JA, Weissman JS, Arkin AP, Lim WA (2013) Repurposing CRISPR as an RNA-Guided...

...the hopes and concerns of these communities. This kind of engagement and two-way dialogue is crucial if...advising writers and directors in Hollywood; and working directly with community organizations, libraries...leadership and communication experience. These included taking extra coursework and attending seminars in bioethics...writing science articles for a lay readership, and working with middle and high school students. One of these...organizations was pgEd. After obtaining my PhD and working in digital academic publishing for a year, I’ve...and in helping you decide whether you enjoy this kind of work. It is also beneficial to hone your non-...

...deposits per month!), manual transformations were taking a lot of time. Enter the Biomek FXP! We wanted ...improve our transformation protocol and, after looking at a few options, we decided on Beckman Coulter...protocol on the Biomek FxP is pretty simple. By clicking a few buttons you can quickly tell the Biomek ...pick up 8 tips and position the tips indepently making it easy to spread out your samples. This can be...scientists share plasmids across the globe, we’re working on ways to send plasmids as DNA as opposed to in...it suits our plasmid prepping needs perfectly. Working with Qiagen, we developed a protocol to run up ...

... (and Addgene depositor) Craig Mello. Taking his groundbreaking work in RNAi one step further, his lab...the majority of attendees were scientists still working at the bench. It was a fantastic opportunity to...cell membrane. His recent work shows that they are making fairly remarkable progress on establishing a system...the drug resistantance genes in the bacteria thus making the bacteria sensitive to known antibotics. These... CRISPR resources Read Joanne Kamens' Post on Networking at Scientific Conferences  ...

... gRNA using restriction enzyme BsaI. If you’re working in Drosophila, a two-gRNA expressing plasmid is...outside of their recognition sequence, creating flanking overhangs. These overhangs can be customized to...vectors each contain Type IIS restriction sites flanking the promoter-gRNA construct, but with different...with the appropriate Type IIS enzyme, the unique flanking overhang sequences can link together to allow ...constructs with multiple promoter-gRNA cassettes, making them advantageous for small capacity vectors like... mammalian option described below, plasmids for making polycistronic gRNAs are also available from the... tyr gRNA, which causes hypopigmentation, thus marking cells that have undergone genomic modification....

...between skin and muscle 25–27 < 2–3 mL (~1 mL per site) Beginner Intradermal Anesthesia Between skin layers...Subcutaneous (SQ or SC) injections are given between the skin and the muscles. The most common injection sites... (ID) injections occur between the layers of the skin. While this could theoretically be performed anywhere...shaved first. Restraint: anesthesia Location: between skin layers Gauge: 26 Volume: < 0.05 ml Skill required...cringe or your experiment will allow you to avoid poking the mice, these “no needles” techniques may come...