...’ve gained experience working with similar protocols, and look - they’re seeking someone who has, to a... a specific field. While searching for jobs and making connections, avoid falling into an identity trap...retrieved your lucky socks, persevered through a nerve wracking commute complete with a disabled train, and now...this job was made for you. They must have been looking at your resume while they wrote the job description...side of an issue. Maybe you also showed critical thinking skills by dissecting their problem and piecing...organizational skills. If you are responsible for working with lots of students, managing inventories in ...pretty much any other workplace. To get your mind working, take a look at the short list of transferrable...

...because you are not (yet) asking for a job or a big favor - you are generally asking for some advice and guidance...researchers do learn MANY transferable skills while working in the lab and by offering advice on areas where...many methods and machines one team can learn so linking together expert teams can push a scientific project...graduate studies at Karolinska Institutet I started working with other groups. First within my department where...meet collaborators  for lunch, make a point of talking to them at a seminars, or even set up regular meetings...much more effective than simply sending a message asking how things are going. Science careers where the...studies in regard to reaching out to new people, checking in regularly, and maintaining useful notes that...

...authors’ compared nicking the top strand first followed by the bottom strand versus nicking the bottom strand...position. The “sloppy” PCR reaction is created by making some or all of the following modifications: 1) ...packaged into phage and then extracted. Besides taking a couple days to generate the phage-derived uracil...ready to be tested in a functional screen of your liking. This method is similar to site-directed mutagenesis...with a twist since it relies on the sequential nicking, degradation, and PCR-generation of a mutant version...Supplementary Note 1 in Wrenbeck et al for details on how nicking was optimized. How should I design my mutagenic...gene of interest has a BbvCI site(s). If so, the nicking mutagenesis protocol will still work if the orientation...

...step by step instructions on creating a budget, making decisions based on that budget, and negotiating...timeline Start with your thesis in mind. Imagine the kind of data your dream discovery will need, and how ...should reflect salaries in the region you’ll be working. At this point, your budget bottom line costs may...position. This is important to ensure that you’re looking at institutes that have the means to support your...values. All too often, we see early investigators making unnecessary short term sacrifices that hurt their...dependent on a pathology microtome/cryostat, then making a careful decision on the condition and availability... and limitations. Everything is negotiable. Try asking for discounts, collaborations, access to internal...

...sgRNA design is not possible without relying on some kind of pre-analysis. At the end of 2014, I began developing...contents spill to the ground. There is something akin to a frenzy as the crowd tries to weigh and determine...each item changes from what the reviewers so painstakingly assessed to something a bit more representative...community as a whole has yet to settle on any true kind of value. I’ve dubbed this the “Piñata Effect”, ...question is then, should we use all tools as some kind of a meta/Frankenstein app? Or perhaps there is ...2013 about 11 tools have been added every year.   Breaking down the barriers: My view on current sgRNA design...released with advanced functionality, it seems these kinds of features are delegated to other more suitable...

...affinity and slower kinetics iGluSnFR3 v857: decreased affinity and faster kinetics which are illustrated... it is selectively activated by glutamate, and tweaking it to improve its responsiveness, they showed ...Improve sensor’s photophysical properties such as kinetics, ∆F/F0, SNR, dynamic range, photostability, quantum...v82) and lime green (v857) variants differing in kinetics, affinity, and ∆F/F0. Introduce the two variants..., GPI, and SGZ, to improve neuronal membrane trafficking and localization. Improved photophysical properties...al…, Supplementary Figure 2.   Improved trafficking and localization through different C-terminal...“Glutamate indicators with improved activation kinetics and localiation for imaging synaptic transmission...

...Scientists are known for asking "What? and "How?" , but they are not always known for asking "Should we?". Scientists...discussed his work with Parkinson's Disease; Hideyuki Okano presented ground-breaking work on spinal cord ...primary Addgene contact for many years, I've been looking forward to seeing how the sharing of his OSKM factors...clinical trials for ALS.   I have an uncle with Parkinson's, a half-brother who is a quadriplegic due to ...techniques are neither fast nor inexpensive.  After talking to numerous poster presenters, it was gratifying... Eric J. Perkins...

...last for several days.” Working with CRISPR Recently Doench has been working on developing CRISPR pooled...not, well, your grade is lower.' ... When you are waking up on a Saturday at like 7 am to go check your ...” Doench has spent most of his research career working on miRNAs and RNAi. He first heard about RNAi at...technology development alongside it.” Doench starting working with David Root in the then RNAi Platform, which...investigate the human genome. Doench says he enjoys working on technology development under the Platform model... but work in true collaboration with scientists asking biological questions. Doench loves that he still... “very much in line with the overall mission of making science easier and more transparent. That’s largely...

...consider when seeking your dream lab. The principal investigator's point of view If you are looking to join...decision. A science trainee will spend 4-7 years working for one person and with a group who all strive ...join a lab, you may be wondering “What is this PI looking for in a new researcher?,” “How do I convince the...typically go through a rotation, which reveals their working habits, personality, and gives them the opportunity...style to spur you to succeed or you might enjoy working closely with others to accomplish your goals. You... You must understand your own style before making this choice.  How big is the lab? While larger labs ... Dr. George Church (Harvard Medical School) for taking the time to speak with us about adding new scientists...

...improve resolution, use a stacking gel on top of the resolving gel. A stacking gel typically has a different...buffer, blocking the membrane  from binding to the antibodies during the stain. The ideal blocking buffer...frequently include suggestions for working concentrations, blocking buffers, and incubation times. If ...interested in making your own knockout line for validation, please view this tutorial from Stuart Orkin's and Daniel...of the proteins in a sample migrate through the stacking gel at the same pace and enter the resolving gel...

...by your library. After blocking for 1 h at room temperature with gentle shaking, gently wick off excess...display little to no homology to anything else, making it difficult to quickly identify possible gene ... overexpression in E. coli, such as pET30a. If working with DNA that is refractory to enzymatic manipulation...the T7 polymerase and a few proteases knocked out making them particularly useful for protein production...harvested dose of cells (preferably in log phase) in blocking solution (around 108 CFU/mL is a good place to...start) and add your membranes to the solution, shaking gently for 1 h at room temperature. This will allow...onto nitrocellulose and carry out the steps from blocking the membrane to probing with cells in order to...

...protein-peptide locking, Mark Howarth’s lab has developed a new tool for peptide-peptide locking. The new technology...light-activated receptor tyrosine kinases Receptor tyrosine kinases (RTKs) are a large class of cell-surface...informational videos, download protocols, and more! Looking for background information on CRISPR technology...Easy-to-use tools to monitor these features were lacking until Roberta Gottlieb and Zhen Yan labs developed...described as “knocksideways” (a British idiom for “taking by surprise”) by Margaret Robinson’s lab, and more...mitochondrial membrane. Sequestration is accomplished by knocking down your endogenous protein of interest and co-expressing... blue and green light-excitable and has faster kinetics than those of other channelrhodopsins. CnChR1,...

... from Leah Bury of Microscopic Art. I started thinking about this problem when I recently attended the...the transcription of a gene a megabase away, or looking at the histone methylation status at a double-strand...double-strand break that needs repairing. They may be looking at chromatin at the level of nucleosomes, loops... other areas of biological research. If you’re looking for resources that will help you study how chromatin...with Addgene, but you’ll find many great tools by clicking on the links below and visiting their associated...Sharon Dent Archa Fox Elaine Fuchs Kai Ge Robert Kingston Thoru Pederson Phil Sharp Julie Ahringer Alexander... listed here. If you don’t see the gene you’re looking for on this list, please consider searching directly...

...information, making it available to everyone, that creates more breakthrough and innovation. With any kind of open...'s operations. We’re looking for those “what could go wrong scenarios” and making sure we have adequate...than just picking up the phone and having a discussion. By only relying on email and never picking up the... more about Addgene and heard that Addgene was looking for new board members. Paratore: Kristen Darby,...goals were and so I decided to join. I’ve been working with Addgene since 2014 so I’ve been able to watch...

...organizations with similar ideas who were also thinking about reproducibility and shared similar desires...Fortunately, the eLife ambassadors group was already thinking about ways to tackle irreproducibility. Benjamin...organizational skills. Meanwhile, Addgene was also thinking about ways to address irreproducibility and thanks... post-workshop surveys, recording results, and tracking material downloads. This was particularly helpful...community. Lessons learned Instead of continuously talking about a problem and potential solutions, take actionable..., but how do you get them to connect? Through networking of course! Thanks to individuals in the group...right time. Be inclusive and seek solutions by working with communities, not for them - Another strength...

...aerosols that are generated (while talking or pipetting) during working in the laminar airflow are also ...providing proper training and guidance to the people working in the lab facility. This can be done by training...talk, laugh, sneeze or cough. Wear gloves while working and discard them after use. Human contact can spread... spread contamination when you are working without gloves or masks. Make sure to change the gloves and...inside the laminar airflow before and while you are working. Perform routine checks for mycoplasma contamination... should be cleaned regularly and dampness of any kind should be avoided. Distilled water provided to maintain...because you are not the first person to face this kind of problem. Educate others when you can and help...

...tool of choice. Nicking sgRNAs PE3 and PE5: These tools use an sgRNA to guide nicking of the unedited...editors bound to sgRNAs only drive nicking (right). Start with nicking sgRNAs ~50 bp from the prime editing... Figure 7: Nicking sgRNAs can be designed to target edited DNA, so that nicking only occurs after... prevents the Cas9 nickase from re-binding and nicking the newly synthesized strand before heteroduplex...BioRender.com. PE3b and PE5b: In this approach, the nicking sgRNA is designed to bind only after the edit is...

... to worm motion. Working on the Cat Microbiome Project Another project we are working with is the cat ...computer scientists, engineers, and biologists working together on a variety of projects including the...cat microbiome project. This is a project looking at the makeup of the bacterial species in the gut of the...expensive to make it practical - but we have members looking at how to re-use it as a platform for monitoring...makerspace somewhere near you. The place to start looking for your local space is the Hackerspace website...Victoria. After a decade at Microsoft Research working on speech recognition, he returned to school to...manufacturing tools – a 3D printer, a laser cutter, woodworking tools, and a metalshop. After two summers in ...

...provide constructive criticism, and ask thought-provoking questions. For example, if you’re a virologist...help build your presentation. For example, when thinking about discussing a figure, jot down the question...poster where the presenter spends the entire time talking to the poster, as opposed to engaging with you?...Even the most prepared of us can find ourselves talking to our posters, so if you find yourself in this...let them know while you appreciate the thought-provoking question, you are unable to answer it at this ...give the “elevator pitch,” of his/her project – kindly let the presenter know you are very interested ...interested in viewing which has drawn a large crowd, making it difficult to see the actual poster or interact...

...while still working towards your degree. For me, experimenting with the career and networking opportunities...workshops and networking events. I took advantage of as many of these resources as I could. Networking and learning... I liked – well, I was used to experiments not working, so just like in lab, I sucked it up and moved ... like a great way to share some of the research taking place at our university with the community. I even...and David Shiffman @WhySharksMatter and see what kind of content they share and produce and how they interact...wanted to do. A few weeks into volunteering, I was talking to a little girl about fossils. After we talked...