...monitoring the first round of translation or by tracking the translation of a single mRNA over time. Both...translation. Single molecule continuous translation tracking TRICK allows you to see the first round of translation...translation, but it doesn’t allow continued tracking of a given mRNA because NLS-PCP-GFP is removed by ...first ribosome that translates the mRNA. To permit tracking over time, Yan et al. devised a system that labels...keeps it in a single field of view for constant tracking. You can then directly monitor multiple rounds...

...Professor at University of Barcelona. The challenge: Making courses on genetic manipulation more hands on While...meeting with some other professors, I went home thinking about how to solve this problem. I didn’t want...your strategy. I realized that similar types of thinking go into developing gene therapy strategies for...noting any gaps in their knowledge. Practically speaking the format I decided on was to have two teams ...skills like team building, collaborative critical thinking, and communication among others. They additionally...

...much of the primary data one might require when making such a decision remains spread across literature...With thousands of scientists routinely using and thinking about FPs, it would be great to have a centralized...building advanced multi-parametric search queries.  Looking for a bright, monomeric FP published in the last...suggestions. If you’re not sure exactly what you’re looking for, the Table of Fluorescent Protein Properties...that set of proteins.  For computer-savvy users looking to incorporate FPbase data in their own projects...

... in Your Working Day" video from Science Media Savvy At this point, you might be be thinking social media...first blog post out of boredom. I had just started working in a new lab and I was the only person in the lab...first post so I signed up to write a second. While working on this post, I was offered and accepted a position...on the Addgene blog Read about online social networking for scientists Learn why scientists should be...

...can leave them at room temperature when you’re working with them, but avoid freeze-thaw cycles, as they...bind to amine groups present in some conjugates, blocking the conjugation reaction. Many commercial antibodies...for your ELISA and Western, I’d highly recommend sticking with what works.  To balance this out, however...in your machine before you make your purchase. Picking a single fluorophore for an experiment is easy,...accurate read and reduce needed compensation.    Working with antibodies can get a bit complicated, but ...

...toolbox openUC2 are trying to overcome this hurdle by taking these limitations into account in the development...software (left) vs. open hardware (right).    Taking this all together, the essential ingredients of...newly developed techniques are the key enablers for making assemblies open-source. The sharing of digital ...produced quickly with the printer, act here as a kind of "teleporter," since a part developed in Germany...way of communicating in science brings stronger networking across laboratory and disciplinary boundaries...

...responsibilities. After my second postdoc, I realized that working in the wet lab wasn’t the right path for me. I ...style of a new, junior team member who had values, working styles, and skills that were quite different from...from them. I started at my previous company by taking on responsibilities such as mentoring a junior ...Beyond Academia: Stories and Strategies for PhDs Making the Leap to Industry." He successfully transitioned...Resources  Resources for aspiring managers Books: The Making of a Manager by Julie Zhuo Radical Candor by Kim...

...protein AraC binds O and I1 sites upstream of pBad, blocking transcription. The addition of arabinose causes... cAMP and represses pBad, decreasing promoter leakiness.   Other examples of chemically induced promoters... be compatible with your experimental system. Leakiness If you’re expressing a gene that may be toxic,...inducible promoters are also known for their low leakiness. Inducibility level How much do you want to induce... inducible promoters can vary a lot. If you’re looking for high inducibility, the Tet-On system may be...

...channel just for asking people if they can write or review things and another for asking specific science...depositors and customers are asking. So how can you get involved? Keep asking your burning questions and...review to catch those pesky typos, and hit publish. Making content is an iterative process. We are constantly...

... account, you can begin the deposit process by clicking "Submit Online Deposit" (for online submission... menu: Figure 1: Begin the deposit process by clicking "Submit Online Deposit" or "Download Deposit Spreadsheet...  Figure 4: Add plasmids to the article by clicking “Add Plasmid to this Article” (left) and entering...button and jump around to the different pages by clicking on the appropriate boxes at the top of the page...the plasmid to copy from the drop-down menu and clicking the white "Copy" button. *Note: The "Copy" button...Submit Deposit" button to complete the data entry. Clicking the "Submit Deposit" button will allow you to ...

... the mice to increase their nose-poking behavior. Strong nose-poking behavior is exactly what was seen...a channelrhodopsin, such as cell localization, kinetics, and spectral properties, must be optimized simultaneously...photocurrent strength, wavelength sensitivity, and off-kinetics.   This model was then used to predict which channelrhodopsins...photocurrent amplitude, wavelength sensitivity, and off-kinetics correlated with the model’s predictions.  Three...

...on everything, including your skin. Here are some general tips for working with RNA, even if you are using...basics of isolating RNA without a kit. Tips for working with RNA (whether you are using a kit or not) Although..., 2006).  View our protocol for RNA extraction! Making the simple protocol even more foolproof (still ...still without a kit!) As mentioned above, working with RNA requires keeping your samples cold until homogenization...

...digest/ligate methods and picking the right one can save you time if you’re working at speed or money if budget...you can explore to maximize the use of both when making plasmids. Use repositories Repositories such as...free. Plasmids ship out 2-3 days after ordering, making it a toss-up as to whether it’s faster to order...experiment. Make your work “high-throughput” Instead of making one plasmid at a time, try constructing several...

... unique cutting features and evolved PAM sites, making Cas9 an adaptable choice for many genome engineering...same mechanism as traditional spCas9. If you are looking to modify RNA, Cas9 just won’t cut it – literally...editing. Fast facts – PAM requirement: protospacer flanking sequence – A, U, or C. Best for: targeting single...use. Fast facts – PAM requirement: protospacer flanking sequence – none. Best for: targeting single stranded...single base pair edits, these enzymes are worth taking a look at. Choosing the right Cas can really put...

... it’s monumentally difficult without software. Tracking lab procedures also gives you access to hard data...repeated. We also now have a good interface to begin tracking analytics data for lab procedures. With this data...are distinct from their contents. You should be tracking both, you should easily be able to move contents...less time getting up and running and more time working on interesting biotechnology code.There are valid...reasons to choose a non-Python stack, but if you're working with scientists it's unlikely they will have exposure...

... There are currently 17 sites around the USA undertaking BEST programs, including local schools in Boston...contributing to the NIH’s Biomedical Research Workforce Working Group. Along with her participation in this panel...Pubmed Central PMCID: PMC3787295. 6. Marder, Eve. "Looking out for future scientists." eLife 3 (2014): e04901...PMC4185419. 7. NIH 2012. Biomedical research workforce working group report. http://acd.od.nih.gov/biomedical_...in Science in Our Career Posts! Get Advice on Networking at Scientific Meetings from Addgene Executive...

...edit and homology to the genomic DNA locus. After nicking by Cas9, the homologous pegRNA sequence hybridizes..., as it can edit near to or far from PAM sites, making it less constrained by PAM availability than other...previously used when they developed base editing. By nicking the non-edited strand, they can cause the cell ...efficiencies 2-3-fold. However, the process of double nicking increases indel formation slightly. Designing the...the edited strand, as in the PE3b system, guides nicking of the unedited strand only after editing has occurred... ✓ ✓ ✓   ✓ Second sgRNA to guide nicking of the unedited strand Favor the edited strand...

...include a lot more. Font size: Making your poster readable While we are talking about font, remember this:...important to reflect on these impressions when thinking about crafting your own poster. What sets a scientific...that it will display your data most prominently, making it the star of your presentation. Portrait. This...presented some standard formatting guidelines for making a scientific poster, you shouldn’t hesitate to ...

...understand the risks of the specific virus you’re working with before you work with it. This will help you...required in BSL-1 and required in BSL-2 labs. When working with lentivirus, many organizations require or ...can quickly be removed and disposed of without risking exposure. You can request disposable gowns from...researchers choose to wear eye protection when they are working in a biosafety cabinet, even if it is not required...used for BSL-2 waste, and of course, if you are working with a substance in a biosafety cabinet, it needs...

...with the NPA where I gained invaluable experience working on and later chairing the Advocacy Committee. I... I’m now furthering my leadership experience working at the Association for Research in Vision and Ophthalmology...website. By volunteering, you can gain experience working with other people mostly online and on monthly ...resulted in actionable tasks and I made a point of sticking to deadlines and performing the work I agreed ...members changed, I had to learn how to develop new working relationships with these individuals who often ...