...unless the source of culture is irreplaceable or highly expensive). Detecting mycoplasma contamination ...induces a noticeable color change. This method has high sensitivity but low specificity (Degeling, 2012)...required in this process because result interpretation might be difficult. The low density of mycoplasmas, contamination...media as much as possible and the ones that are tightly packed with seals. Use disposable pipettes to avoid...exposure of the cell culture to air and make sure to tighten the flask cap before placing them in the incubators...

...decades of my life following a very straightforward conveyor belt from high school to college to graduate school...Veterans Health Administration, I could never get one straight answer about what science policy even is! In this... “little p” policy.  “Big P” policy is what you might typically think of. It occurs in a government setting... challenging if you are trying to apply for an outright job in a field where you have no specific scientific...

...and four chaperones from Denis-Yarmouth Regional High School visited our labs in Cambridge, MA to learn... Outreach Scientist Jessica Welch showed some highlights of his travels from 2015, including a great European...Website Learn more about Dennis-Yarmouth Regional High School Resources at Addgene  Read more Inside ...

...working with AAV or lentivirus in your research, you might want to use one of our virus backgrounds. Addgene...backgrounds for you as well! If you use FRET, you might be interested in our fluorescent protein background....           Did you get the joke? If not, you might want to check out Addgene’s Fluorescent Proteins...background.     We hope these science backgrounds will brighten up your virtual meetings!  ...

...It’s well known that exercise can help fight stress and depression, and now researchers have reported...including unpredictable loud noises and flashing lights - that sent average, control mice into depression...know about you, but I think I could use a good run right about now. And, if you are looking to get your hands...for studies in your own lab, you’ve come to the right place. Get started with PGC-1 plasmids from Addgene...

...Directors brings human resources experience and insight to the now 13 member board at Addgene. Let’s meet...hope Addgene will accomplish in the near future? Right now it’s a matter of maneuvering through the impact...volunteer appointments included the Belmont Human Rights Commission and mediator for the Massachusetts small... dedicated to volunteerism and ran the Mayor’s Midnight Sun Marathon in Anchorage, for the Leukemia Society...

...to the field. Fortunately, our bodies have been fighting cancer for far longer than we’ve been studying...natural abilities, we’re gaining new ground in the fight against cancer in its many forms. The dichotomous...cancer stops growing, and the immune system stops fighting. Such quiescent cancer may rear its head once ...infection can further divert the immune system from fighting cancer. Surgical removal causes increases in tumor... a sequence of immune reactions that aid in the fight against cancer. Sipuleucel-T by the Dendreon Corporation...

...instance, you might use it as part of a flex switch you’d like to work with in neurons or you might use it in...Cre-controlled gene of interest makes pmSyn-BFP-Cre highly modular. Further uses of pmSyn1-EBFP-Cre Of course...brain. There are many possibilities enabled by this highly modular AAV - that’s probably why it’s so popular...

...bigly. But such a project should not be undertaken lightly. Whether executed in arrayed or pooled format there...are just as critical for success. Sometimes, you might have a few positive control genes on which to test...execute genome-wide perturbations. So, while some might argue that the only way to do something is to do...still offering the potential for true biological insight. Pooled and individual plasmid formats A first ...

...group? The Ellis Lab self-organizes cinema nights, game nights, and trips to eat out together. “We also ... celebrate your lab’s achievements? Tom Ellis highlights the importance of acknowledging his teams successes...

...? If so, you might want to find additional sources of wisdom. While somewhat lighthearted, the article...article does highlight key characteristics of a good mentor: Yoda image courtesy of jorenl/Vecteezy.com...

...share? And how they are connected? Here are the highlights written as 8 tweetable facts. Note: The images...most Twitter users. Biologists rank #8. Figure 1: Highly Represented Scientific Disciplines on Twitter. ...disciplines: social (left), life (center), and physical (right). How do you use Twitter as a scientist? If you ...

...them share their reagents, we knew our role in fighting the pandemic could be even more impactful by reaching...scientists working in a for-profit setting.  In light of the need for basic research during the current...University of Iowa University of Minnesota See highlights from the COVID-19 collection Partnership with...

...Agrobacterium, along with a selectable marker. Left and right T-DNA borders specify the region of DNA that will...codon-optimizing the Agrobacterium ORI, they achieved much higher DNA yield and transformation efficiency in both...that gene expression from the M24 promoter is much higher than that from the commonly used 35S promoter. ...transfer. The M24 promoter provides constitutive, high-level gene expression and is functional in a variety...

...been in space is pretty small). Therefore, I was delighted to be invited to the ceremony for surely what ...non-scientist from Bloomberg news. Given the topic it might have been better to have a scientist up there, but... viruses to give information to the computer to fight the known viruses. Bacteria have figured out how... virus sequence to enable them to recognize and fight it forever. He emphasized the potential of this ...

...toxic to both lines, it probably has either high toxicity or high off target effects, both of which make it... druggable target). Diseases with defined, straightforward causes (a specific mutation in a known gene...targets by CRISPR can accelerate the drug pipeline. High-throughput CRISPR screens can be used to inhibit...

...research". The family entered the biomedical field’s spotlight after researchers discovered heavy-chain only ...reading frame Ability to access unique epitopes with high affinity, due to a greater ability to target concave...cellular functions, like in this early example we highlighted back in 2015, where sdAbs were used to create...

...Gates’ Foundation is involved with was one of the highlights of the conference. It was inspiring to see how...trusty spray bottles! We also raffled off one of our highly sought-after "nice genes…" tshirts, and the lucky...Fluorescence Labeling Plasmids from the Barbier lab). Bright and early on Saturday morning, John Doench’s opening...

...skills to get the work done efficiently and with high quality. Their work involves coordination and can...t mean going rogue, pointing out problems or highlighting criticisms. To be given the opportunity to take... by Peter Fuda and Richard Badham Leadership Insights for Engineers, Scientists and Computer Professionals...

...planning to transform right away OR you can store the reaction at -20C overnight. 4. Transform TOPO Cloning...and requires no post-PCR procedures. Sounds easy right? The technique relies on the basic ability of complementary...efficiency).  5. Select and Analyze 10 White or Light Blue Colonies: You can confirm the presence of your...

...neuroscience with ever more efficient tools for light mediated neuronal control and we are expanding our...target photostimulation of axons and dendrites on neighboring neurons. We have also packaged hChR2(H134R) plasmids...apparent dopamine affinity, with 113050 offering high apparent affinity, and 113049 offering medium apparent...glutamate levels. iGluSnFR.A184S, a variant with higher affinity to glutamate, has been packaged into AAVrg...

...isotype converted, or chimeric, antibodies may be the right solution for you. Read on to learn more. What is...is an isotype? Antibodies are composed of light chains and heavy chains made up of variable regions and...097037-0.00017-8     Morelock, M. M., Rothlein, R., Bright, S. M., Robinson, M. K., Graham, E. T., Sabo, J...Polyclonal Antibodies Antibodies 101: Selecting the Right Antibody  ...

...Bundling up for winter? The weather outside might be frightful, but browsing the 49 new preps available ...absolutely delightful. Grab a hot beverage and a blanket, then scroll on for some wintery delights!    Table...

...way? What ecosystem makes your heart sing with delight? From developmental biology to developing a career...example, how to use new technologies to reach the right audiences and how open access affects financial ...of scholarly publishing. And see for example the Eighth International Congress on Peer Review and Scientific...interested in the science communications world, you might consider an internship or a short program to build...

... Addgenie Julian Taylor-Parker explains why you might want to use a specialized E. coli strain for protein... to an entirely new career path, a career coach might be able to help you. Head Addgenie Joanne Kamens...genome, you need to check whether you’ve made the right one. In this post, Addgenie Melina Fan guides us...guides us through some common controls and why you might need them. Happy Reading!   Resources on the Addgene...

...This guest blog post was contributed by Seán Mac Fhearraigh, PhD, of www.phdadvice.com. I sold out, well ...Like a considerable number of researchers, I had high hopes of securing tenure in a British or Irish University...interviewed for 1 hour so having a broad enough insight was going to help me push both my scientific and...death assays. You can find Seán on twitter @Macfhearraigh.    Read more posts about Scientific Career ...

...amplification and verification processes we use to ensure high-quality library distribution. You can also use these...amplification 101 Library amplification seems straightforward: E. coli bacteria are transformed with the ...a typical library, Addgene usually does four to eight electroporation reactions using a total of ~400ng.... Analyzing the reads is, again, seemingly straightforward – you need to count how many times each of ...

...involves measuring the absorbance/transmission of light through a liquid to determine the concentration ...liquid. Molecules absorb different wavelengths of light to different degrees and many molecules have a specific...method takes more time to set up in the lab, you might not have to do the calculation yourself as many ...disadvantages of each method and when one method might be more suited than another. Additional resources...

...to 3-loci MPE with similar high efficiencies, which is scalable for a higher number of MPE. Therapeutic...leader sequences of a given tRNA to achieve the highest efficiencies of multiplex editing at each site....Cas9-dependent off-target editing and do not cause higher Cas9-independent off-target editing in comparison...

...sequences are from AAV2 because they demonstrate high promoter activity (Earley et al. 2020), but non-...downstream AAV applications.  Tips & FAQs Due to their high GC content and hairpin structures, ITRs are prone...rapidly replicate in a growth culture and result in a high percentage of plasmid DNA with mutated ITR sequences...Next-generation sequencing and Sanger sequencing might fail to correctly read through ITR sequences. GC...

...is not just a question of being honest and straightforward, it’s an important part of being a “good scientific...work to figure them out. But, when they come to light, it’s clear that what the researchers thought was...still missing is a clear alignment of incentives. Right now, researchers are not strongly rewarded for being.... But it ought to be possible, and if we get it right, everyone in the scientific enterprise will benefit...

...protocol, get a result and hope it’s close to the right answer because if it’s not, well, your grade is ...the data look like... when it looks good, that’s a high that can last for several days.” Working with CRISPR...Doench thinks that the future of CRISPR will be in high throughput functional characterization of specific...plasmids, figure out what they do and what they might be able to do and to organize it and catalogue it...

...have purified plasmid DNA, this method can be done right in your lab in less than a day. Diagnostic restriction...sequence if necessary. The example plasmid on the right has a total size of 7.3kb, including a 1.2 kb insert...recognition sequence. Most enzyme manufacturers make High Fidelity versions of the endonucleases and/or supply...you to visualize the DNA under ultraviolet (UV) light. Don’t forget to add loading buffer to your digest...

...succeed in communicating. Even the most rigorous, insightful, and novel scientific research will be wasted...your past work has prepared you to contribute at a high level to their team.  When you're writing a grant...anyone else in the world to do it.  In the lab, it might mean persuasively articulating why the results you've...to be reviewed by interdisciplinary panels.  And highly successful scientists need to communicate with ...

...instrumental in helping us establish consistent, high-quality protocols for lentiviral production. Bryan...Janelia Research Campus have provided valuable insight into AAV production. Finally, we are blessed to...surveyed hundreds of scientists about various ways we might expand Addgene’s services. There was an overwhelmingly...helps scientists and allows us to maintain Addgene’s high quality standards?” Under the leadership of Addgene...

...labeling tubes never takes longer than 5 minutes, right?) As a result, I’d always end up stressed and rushing...add in personalized steps like “check drawers for light-sensitive reagents” or “re-sterilize now.” Have ...Moving your reagents from your left side to your right as you use them can help you remember what's been... Fig. 2: Blugene moves reagents from left to right as they use them.    Prevent Interruptions If...

...Viral integration of a construct is also reliably higher than traditional transfection and random genomic...transfection method. This is because viral systems have a high infection frequency and the viral titer (the number...responses in addition to antibody response 2) very high immunogenicity 3) and long-lasting immune response...treatment and gene therapy have also been in the limelight. The FDA approved the first oncolytic viral therapy...

...BioBuilder Educational Foundation in partnership with high school teachers to boost interest and understanding...what started in Cambridge with a handful of local high school teachers receiving training modules in synthetic...fall:  The BioBuilderClub BioBuilderClub engages high school teams around the world who combine engineering...and STEM-loving students include The BioBuilder High School Apprenticeship, preparing local (Massachusetts...

...disease. CRISPR has made genomic SDM relatively straightforward, but plasmid-based SDM has lagged behind. While...well, but poor template digestion can result in a high number of background colonies carrying the unmodified...plasmid is not maintained. Recombination efficiency is high enough to negate the need for selection usually ...introduce mutations. Using commercially available high-fidelity polymerases such as KOD and Phusion, which...

...Mendenhall: The benefits are two-fold: CRISPR/Cas9 offers high efficiency of inducing double strand breaks and ...weeks. CRISPR saves you a couple of months. It’s higher throughput and faster. It’s also easy to run things...transcription factors, but there are many reasons you might want a system like this. It’s actually surprising...with this antibody problem. We thought it was the right thing to do to put these out there. It’s a simple...

...interested in following? Someone you admire for their insight and way of expressing it in meetings? The more ...advice. Your family and friends know you well and might have good ideas for who you should connect with....cautious and make sure to choose one that comes highly recommended from someone you trust. A good coach...compliment to ask someone to share their wisdom and they might just not have the time. Thank them anyway and move...

...result in any cells, or all the colonies screened might not contain the correct plasmid. There’s a lot that... 24 bp with the template and if the sequence has high GC or AT content, increase the overlapping region...Full lawn of cells. The antibiotic in your plate might not work. Check the plates! Streak on them an E....t get any positive clone after these steps, you might want to reconsider your approach, and try a different...

...concentrations. Higher IPTG concentration leads to higher RepA expression which results in high plasmid copy...repressor because TALEs can be programmed to bind tightly to arbitrary DNA sequences and have been shown ...

...interactions by cross-linking RNAs and proteins with UV light, followed by an antibody pulldown of a protein of...for Affinity Purification (CRAPome), to identify highly biotinylated background proteins reported by other...and identified QKI, an RNA-binding protein that’s highly expressed in neural progenitor cells. Knock-down...Response Element (IRE) RNA motif of the ferritin light chain RNA transcript. IRE motifs are bound by IRE...

... is weaker than reporter staining. Picking the right reporter for your RANBody While all three reporters... fluorescently labeled tyramide molecule into a highly reactive radical, which is then covalently linked...antigens. Spaghetti Monster Spaghetti Monsters are highly antigenic tags that have 10 HA or 10 MYC tags built... plasmids from Penn State Need help picking the right fluorescence microscopy technique to use? Check ...

...feels she was lucky to have been in the right place at the right time when she started her scicomm career...“There were jobs that I didn’t seem to have the right skills for in terms of PR types of things or communications...leverage unpaid work for paid work: “If you use it right, it could help you get your name out for stuff that...

...neutral pH compartment, ­­like the cytosol, to the higher pH of the vacuole. Read on to learn more about ...enzyme. Pho8 phosphatase activity or the molecular weight shift of Pho8 from its uncleaved to cleaved form... dual color-emission biosensor named after the brightly-coloured Australian parrot. It’s comprised of ...Oligomerization Seeing Red: Simple GFP Photoconversion Light Sheet Fluorescence Microscopy Additional Resources...

...cyan, or magenta light. A gRNA with all 3 distinct hairpin sequences produces white light. Created with ...from red, blue, and green channels overlaid on a bright-field image. Image from Ma et al, 2016. Using... A gRNA with all three hairpins generates white light. All 6 gRNAs can be expressed in a single vector...

... edition of “What’s New in CRISPR” - where we highlight a few of the newest CRISPR plasmids available ...cells. The “V4” version (named CasMINI) had the highest activity for gene activation  and base editing,... not modify the Cas12f1 enzyme. The result is a highly efficient Cas system with very compact genome editors...

...RNA! Double stranded? Well, that’s just for DNA – right? RNA performs almost all of its biological functions...replication process. dsDNA viruses yield dsRNA through a slightly different mechanism thought to be associated with...always around. R-loops are particularly enriched in highly transcribed regions and repetitive regions, such...RNA:DNA hybrids levels are allowed to reach very high or low levels, possibly due to dysregulation of ...