...2015). Meanwhile, Sanger sequencing is laborious, time consuming and cannot be applied to heterogenous ...multiplexing) Requires large cell population, senstive to time since cell fixing HTGTS Not reported Cell-based...sequencing techniques, please see “Methods for Optimizing CRISPR-Cas9 Genome Editing Specificity” by Tycko...Vic E. Myer, and Patrick D. Hsu. "Methods for Optimizing CRISPR-Cas9 Genome Editing Specificity." Molecular...

...synthesized and degraded within the cell, so at any given time, an FP-chimeric protein may be at any one of many...quenching”). The photoswitch can occur multiple times by activating with two distinct wavelengths of light...possible excitation lasers) it is also important to optimize your activation protocol for your particular set...heavily on the same fluorophore being imaged multiple times in order to reconstruct an image. The best PA-FPs...

...as confusing as reading Shakespeare for the first time. The discovery and application of CRISPR-Cas9 kicked...bacterial and archaeal immune systems. During this time, scientists discovered many related immune mechanisms... only recently that researchers began building optimal plasmids to harness the power of Class 1 systems... your goal, experience, hypothesis, and even the time and money available. The breadth and diversity of...

...distributed over 800,000 plasmids since 2004. In that time, our lab has gotten a lot experience in processes...tip #1: Glove trick Putting on lab gloves can sometimes be annoying. To make it easier, try this quick...a single tube and simply flick the tube several times. This will help ensure ensure equal distribution...

...contributed by Kusumika (Kushi) Mukherjee. The ultimate goal in the field of regenerative medicine is ...iPSCs into the cell type of choice. This step is sometimes also referred to as “directed differentiation....39]. In 2000, Ferber et al. showed for the first time that mouse liver cells could be transdifferentiated...Partial rearrangement from one cell type to another Time and resources More (due to additional pluripotency...

...is recognizing the PAM on DNA, the CTD is also sometimes called the PAM-interacting (PI) domain. The guide...al., 2013). As mentioned above, however, Cas9 sometimes tolerates mismatches in the next 8-10 bases after...and the DNA sequence is recognized, it’s finally time to cut! The HNH domain, which cuts the target DNA...Shima, T., Nureki, O., & Uemura, S. (2018). Real‐time observation of flexible domain movements in CRISPR–Cas9...

...Thoughts on his time in Boston and Outro We'd like to thank Dr. Kwapata for taking the time to talk to us...across the globe. Listen here! Podcast Breakdown (Time: Topic) 0:00-1:00 : Intro 1:01-3:48 : Kingdom Kwapata's...

...here at Addgene HQ in Massachusetts that means it’s time for near-freezing temperatures, our first of the...decorating contest! Our #DeckTheLab contest is a time for scientists worldwide to show off their decorating...little more festive this holiday season! It is that time of the year again: miniature #Christmas tree goes...

...graduate student life. Senior graduate students can sometimes be elusive or have a certain mystique that makes...Listen to the interview here! Podcast Breakdown (Time : Topic) 0:00 - 0:28 : Niroshi and Ben describe ...We'd like to thank Niroshi and Ben for taking the time to sit down and talk to us and wish them the best...

...origin. The type of cloning approach you choose ultimately will influence the quantity of DNA that is required...viability, reduces transfection efficiency, and ultimately affects the quality and outcomes of experiments...plasmid DNA, so that you can decide what will be the optimal type of plasmid preparation in terms of quantity...

...meetings, and finding the best way to organize your time. At a conference, you could be honing your science...can't wait for opportunities to fall in your lap. Sometimes, in order to do the thing you love, you have to...  Break the process down into steps and set aside time during the week to tackle each one.  Get the toughest...

...Addgene’s compensation decisions. It was a great time for me to join the committee because I had recently...community? Access to plasmids which gives people more time to focus on their research.  How will your prior...that I’m not working so that I’m available to spend time caring for my mother. For fun, I enjoy pilates, ...

...you need the right experimental materials. Who has time for paperwork? Here’s why a Material Transfer Agreement.... Here’s how Addgene helps: For Providers: One-time Deposit: Simply deposit with Addgene (it’s FREE!...Keeping: Addgene will let you know exactly how many times your materials have been requested and by whom. ...

...technology with the Golden Gate method. For the first time, it was possible to quickly and efficiently assemble... in subsequent rounds, dramatically reducing the time required to generate multi-construct transgenic ... half a year,” in comparison to more tedious and time-consuming traditional cloning methods, Lohmann said...

...are involved. They may feel they are wasting their time. The manager may appear to be too busy to be receptive...team: Solicit feedback regularly and often A one-time event doesn't work to get people sharing.   Frequently... for feedback and input on multiple topics every time you meet.  Seek feedback in multiple formats      ...

...knockout mice, which is time-consuming, as it only changes one gene at a time. CRISPR-based systems enable...functions in different cell types, at different times, and in combination with different genes. Having...

...academic inertia, it’s time to develop systems that give trainees a plan, both for their time in the lab and ...students and post-docs alike are wondering where the time has gone. Are we any closer to graduating, publishing...

...just the “training” portion of our careers. In that time we may have only 2-3 formal supervisors that will...think you can finish that book in 4 weeks!)” “Sometimes a mentor is a sounding board--a good mentor asks... risks A great Mentor is often one that devotes time and energy to thinking about how to be a great Mentor...

...out in a myriad of journals every week, it can sometimes be difficult for great work to gain prominence.... Addgene's "How to cite this plasmid" section Anytime you use a plasmid from the repository, we request... to repeat the modification. This will save both time and money. A quick note -  While it is tempting ...

...accelerate your research. We know you don’t have time to be waiting on hold, let alone waiting multiple...support team available whenever you need them, but sometimes that’s not enough. With your busy schedule, you...features that allow users to help themselves, saving time and effort for everyone involved. We’re adding self...