...Addgene Blog Mentoring for Scientists Series. I have been thinking a lot about Mentoring for over 10 years...group mentoring format could work wonders.  I have now seen the formation of hundreds of mentoring groups...also serve as a great first-time mentoring experience. They make mentoring less mysterious and train the ...useful for future mentoring relationships.   If you would like to start a group mentoring program at your...interested peers and form your own peer mentoring circle. In a Peer Mentoring Group all participants act as both...and what you want to cover during the year. Suggest that each person bring 3 topics and write them on ...involved in Peer Mentoring relationships? Comment below with ideas from your own mentoring group or with ...

...to learn about her mentoring style. Top 7 questions scientists ask me about mentoring In this blog series...the other six mentoring questions in future posts. If you have a question about mentoring that you want... enough?  For many years I’ve been organizing mentoring programs for scientists and doing training to ...fulfilling, productive relationships. I will be sharing what I’ve learned along the way in this ongoing... to the top questions scientists ask me about mentoring. Here they are: What makes a good Mentor? How...? What is this I hear about “group” or “peer” mentoring? How do I ask someone to be my Mentor? I have ...certainly realized the importance of seeking out mentoring as an important aspect to their ongoing success...

...bacterial CRISPR engineering One common way of accomplishing bacterial genome engineering is done with recombineering...combines CRISPR and recombineering-based MAGE (Multiplex Automated Genome Engineering) technology (Ronda...combines recombineering with CRISPR to create a system for scarless, iterative genome engineering (Jiang...promoters in Pseudomonas syringae and Klebsiella oxytoca. Microbiome engineering Scientists have begun to...discovered in bacteria, most CRISPR-based genome engineering has taken place in other organisms. In many bacteria...quirks change the way CRISPR-mediated genome engineering functions in bacteria, but have no fear - plasmids... recombineering, a technique that utilizes phage recombination machinery to promote homologous recombination...

...KTRs are not well suited for monitoring kinase activity that occurs during nuclear envelope breakdown (...important for regulating signal transduction and measuring kinase activity at the single-cell level can aid...signaling activity and cell phenotype. One method for monitoring live single-cell kinase activity is FRET, but...Advantages of KTRs When compared to FRET reporters for measuring kinase activity, KTRs have two key advantages....

...multiplex genome engineering. Image from Sakuma at al. (2015). Multiplex genome engineering demonstrated...system for CRISPR/Cas9-mediated multiplex genome engineering is now available thanks to researchers at Japan...making an all-in-one vector for multiplex genome engineering.” Now, they’ve filled that gap. As Sakuma and...efficient viral vector-mediated multiplex genome engineering. The system now provides an efficient targeting...Sakuma’s protocol. Reference: Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector... multiplex genome engineering...

...database and ordering platform soon to be released. For individuals interested in ordering from ABRC for...Learn how BioBuilder can bring Synthetic Biology into the classroom Engineering the Plant Genome Using ...to us at [email protected] for help setting up an ordering account. Using Arabidopsis in the classroom has...CRISPR/Cas9 pSiM24: Simplifying Plant Genetic Engineering Resources on Addgene.org Find Plant Vectors...

...platform for sharing, annotating, and storing life science protocols. Since we began sharing this protocol...Developing Lab Management Software    Organizing and sharing your updated protocols So that’s great, but what...Plasmids 101 blog series. Want to learn more about sharing your plasmids via Addgene? Check out all the benefits...feature further simplifies lab work if you like to bring your phone in the lab, though, honestly, this still...non-bench work – but is there a sterile, clean way to bring your phone into the lab? Maybe having a dedicated... protocols listed on OWW, including those for preparing your own competent cells. If you’re familiar with...exchange currently boasts over 2,500 protocols covering a variety of bio and biomedical topics. Protocol...

...using glass cloning rings to isolate colonies of antibiotic-resistant cells; during the time required to... to grease, place and fill a dozen cloning rings, the remainder of the colonies on the plate dried out...out and died. The alternative to cloning rings, dilution cloning into 96-well plates, is extremely time...minutes- a far more rapid pace than using cloning rings. A picture of a 60 mm dish ready for picking is ...differ grossly from those isolated using cloning rings in terms of growth rates, morphology or other physiological...

...bad news. The bad news is that some of tubes’ o-rings are, in fact, permeable to CO2 at low temperatures...What we found Not all tubes (or rather, not all O-rings) were permeable to CO2 at low temperatures. We tested...Conclusions and handling recommendations Not all O-rings function optimally at -80°C. For samples that are...or not the dry ice we use to keep viruses frozen during shipment was having an impact on the samples. We... Order Ready to Use Viral Preps from Addgene Measuring the pH of our samples: What we did To measure ...CRIPSR User (By a 1st Time CRISPR User) Tips for Titering Your Lentivirus Preps Additional Resources on...

...: they require switching between a two-ring purine and one-ring pyrimidine base. This is a far more complex... the side groups of DNA bases and leave the core ring structure intact, allowing them to make 4 out of...backbone, making them efficient and precise tools for altering DNA sequences. These first base editors were both...researchers in the Keith Joung lab, who were engineering an ABE to reduce its tendency to make off-target...sites. To explore this further, the team started engineering a CBE to be worse at its original job. They ...glycosylase inhibitor (UGI) components, which inhibit BER during cytosine-to-uracil editing, and instead introduced...GBE editing but not CBE editing, maybe because repairing a GBE-derived abasic site requires more cellular...

...underside of the “umbrella” of A. victoria there is a ring that glows with a faint green light. To investigate...

...accessible and easier to share. As we prepare to ring in the New Year, let's make a toast to open science...those of us at Addgene believe in the power of sharing, and especially in keeping the products of science...

...give to your questions. If one of their answers rings true for you, go ahead and give a similar answer... to the hiring manager and others, if you have their emails. You can use notes you took during your interviews...Fielding questions can be hard during this talk since you’ll be sharing your screen and won’t be able ...2016 and then working four more years in academia. During my job hunt, I quickly noticed one of the biggest... and/or small groups, usually starting with the hiring manager. These interviews lasted for 30 minute ...a list of who will be interviewing you, ask the hiring manager or recruiter for this information. I found...presentation to your audience Start by asking the hiring manager or recruiter what presentation format they...

...a protein based on the affinity of the imidazole ring of histidine for metal ions such as Ni2+ or Co2+...the polyhistidine tag? The polyhistidine tag is a string of 6 or more histidine residues expressed in frame...step to remove the tag in order to prevent it interfering with protein function. In contrast, the polyhistidine...using these more selective substrates along with stringent wash conditions.   Find polyhistidine tag plasmids...Sometimes a tag placed on one end will be blocked during protein folding, in which case you should try tagging...neither placement works, consider purifying under denaturing conditions. Unlike other affinity purification...). In some cases, proteins can be bound under denaturing conditions to expose the tag and then refolded...

... position. Moving one position over in the sugar ring, a popular stabilizing modification is 3’ phosphorothioate...

...Stephen, Andrew G., et al. "Dragging ras back in the ring." Cancer cell 25.3 (2014): 272-281. PubMed PMID:...computer modeling have enabled researchers to begin discovering and designing molecules that are showing some...

...discovered that incorporation of four-membered azetidine rings could substantially enhance the brightness and photostability...

...actually has chimes outside of our break room. A ring of the chimes means something fun is going on and...in a unique environment where, in addition to furthering our mission by hard work, we genuinely enjoy ... that you don’t work for Addgene (though we are hiring!) but to encourage everyone to make an effort to... the power of simple word of mouth - be sure to bring up your event in casual conversation to get people...

...to help students during an interview. When they were instructed to use the three-ring binder as a starting...transferable skills learned while working in the lab and offering advice on areas where you might need some help...creativity is absolutely a transferable skill developed during PhD training. Creativity is required to develop...Whether it’s finding the source of contamination or figuring out why the lab is so cold, we’ve all had to think... need ahead of time and mute your notifications during. (Boyes, 2019) If you’ve taken the time to do this... assure that all participants feel comfortable sharing their ideas? And how do we give people the opportunity... The purpose of this form was to encourage the sharing of ideas, not to give people more work. Identify...

...camera lens without having to physically rotate the rings around the lens. It allowed me to focus on the subject...constraints given. And there were many constraints during the COVID-19 pandemic. The Pandemic Pivot Back...Even Blugene is practicing social distancing and wearing their mask. But while the camera is here, the operator...chair not watching their actors live on set, but peering at a 7-inch wireless monitor. Wireless video technology...control, and extra hand sanitizer.    Cycle 4: Venturing into the Lab Design: The lab is the beating heart...heart of Addgene. It’s why we exist, storing plasmids, and distributing them to labs that need them. The...projects lined up, some that started production during the middle of lockdown, and some that will spin...