...contains a region with a high amount of guanine and cytosine nucleotides, we might need to send it for Sanger...to confirm the sequence of a plasmid and we’ll highlight some of the new features available on our website...our first clue about where the gene of interest might be located. By easily using BLAST to check the translated...repetitive region is present in the plasmid, we might need to perform a restriction digest to confirm ...

...get to where I was. It was really luck of right place, right time, and having a good interview experience...educators, mostly elementary school, middle school, and high school.  I’m the only science person of the bunch...plans was paved with lots of writing, hard work, a slight detour, and a bit of luck. This journey started... editing These teaching experiences definitely weighed in Caitlin’s post-grad plans, but in the end, writing...services. “I was working during the day and editing at night. It was not the easiest 6 months. But it gave me...

...Scientist I/II. Promotions don’t happen overnight though, and it might take longer to advance without a PhD... few questions to help you decide if a master’s might be for you. 1. Are you interested in master’s-...with a PhD, a master’s is less specialized so it might lend itself more readily to a switch. 4. Do you ...the bench. So if you enjoy wielding a pipette, it might make sense to earn a master’s instead of a PhD. ...good fit with your interests and career goals. You might still decide to earn a PhD, but if I had to do grad...

...raging night, With strong heats full, our souls ignite, When all seems lost in the War of Light, Look ...After three kids, I had put on typical middle age weight and “exercise” consisted of taking walks in the...career as Senior Director of Biology, is helping us fight against FOP in two ways. First, Addgene is distributing... their plasmids with Addgene and join us in the fight against FOP. Second, Addgene will be hosting the...                                    -for HOPE burns bright!"   Many thanks to our guest blogger Kurt Swanson...

... your professional brand. It offers colleagues insight into your interests and will determine how you ...Advice from principal investigators Tom Ellis highlights the importance of keeping up-to-date with the...project that aligns with your dreams. He also highlights the importance of combining “new discoveries ...and mentors will help elucidate which project is right for you.   Acknowledgments Thank you to Dr. Tom...

... functions using slightly different Cas proteins and PAM sequences. A total of eight evolutionarily conserved...safeguard against bacteriophages did not come to light until 2007. Initial experiments exposed S. thermophilus...undiscovered viral anti-CRISPR genes. These data highlight an interesting "arms race" between phages and ... to work diligently and creatively to deliver a highly specific, programmable platform well-suited for...

... areas, including one hot spring requiring an overnight hike in. Navigating challenges: collecting and...and processing environmental samples Since the high temperatures and unstable ground around these hot springs...spring. Hiking all day and processing samples by night was fun but also very tiring. It feels a bit strange...gloves at a campsite. And it was exciting – one night while trying to process samples at the picnic table...using microfluidic parallelization to sequence this highly complex hot spring sample. The sample was separated... sequencing, allowing for lower complexity with higher throughput and rigorous assembly. There, they found...

...Fighting with antibodies to produce immunohistochemistry images that are crisp, bright, and lacking in...find high levels of background staining by the secondary, then the following three strategies might help...strategies for mitigating this issue, and we’ve highlighted a few below.  What’s the problem?   First, why...

...well-organized, documented, and easy to find (though we might need to see it to believe it.) As you browse the...epitope that was previously hidden, an antibody might suddenly work that didn't before. This is most often...antibody to bind to a previously hidden epitope (right). Created withBioRender.com.   Biological Processes...it is attached to a cell membrane.  Picking the Right Antibody  It can get even more complex if you’re...configuration. So how do you ensure that you have the right antibody for the protein as it exists in your assay...how the available epitopes in any given protein might change depending on the conditions and the assay...

...very high level of independence, such as museums, YouTube channels and podcasts, science pub nights, and...plumber, a physicist, an ambassador to Spain, and a high school student would all be defined as laypeople...comes to molecular biology.  Some lay people have a high amount of knowledge or interest in STEM subjects...it would be safe to assume that they both have a higher level of science knowledge than average and are...scientist underrepresented minorities in your local high school who are also students in AP biology classes...

...drastic shift won’t happen overnight, but there are a few reasons why you might choose NgAgo over CRISPR...TtAgo-mediated cleavage in mammalian cells, likely due to high temperature requirements. To solve this problem,...preclude future HDR, which could lower HDR efficiency. Higher sensitivity to off-targets More research is needed...other small nuclear DNAs from binding NgAgo, it might make it difficult to use NgAgo to disrupt multiple...Addgene Blog Read About Cas9 Homolog Cpf1 Check Out High Fidelty Cas9 Mutants Browse All Genome Engineering...

...explains, while these Twitch sensors are always bright, they change color as they shift from "off" to ...They identified a toadfish TnC variant which had a high-affinity for calcium binding and developed a version...see tonic action potential firing in neurons and high resolution functional tracking of T lymphocytes,...and acceptor domains. He says, these new colors might now enable observation of the interaction between...Addgene? Check out our Hot Plasmid page, where we highlight some of the popular & newly available plasmids...

... sections of scientific articles (gasp). To my delight, not only was I able to make the edits without ...all about the MOI. In retrospect I used a way too high Cas9 MOI considering that A549 cells are relatively...robust cell line. If your cells are sensitive, it might be more difficult to get them to grow out. The only...presence of the band, you could run another gel that highlights the important results (for example, if you needed...a human cell line with a gene knockout was straightforward, and is a testament to how far genome engineering...David Root and is described in Rational design of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation...

...butterflies in your stomach, the virtual approach might be less stressful. For both conferences, there was...Bachle. (For presenters, there’s also less stage fright!)And depending on the conference, the audience ...can upvote a question and the host will see the highly voted questions,” says Zhang. “Usually the moderator...moderator starts by asking the speaker the highly upvoted question.”If the Q&A session ran out of time, ...for Everyone.  While this type of interaction might not work when you are giving a research talk, there...difficult in the virtual format than in-person. You might miss out on the short chats between sessions or ...

...reporter that has a large dynamic range and high brightness. The Shu Lab from UCSF designed an improved...reporter that has a large dynamic range and high brightness. This reporter achieves approximately 100-fold...immune system, and cell turnover. Apoptosis is a highly controlled process that is triggered by internal...a TEV protease cleavage site. TEV protease is a highly specific protease that is commonly used to cleave...

...match your needs. If YFG is thrillingly new, you might have to make a transgenic fly in order to study ...study it. The process is fairly straightforward. Basically, you place your gene of interest in a plasmid and... trying to overexpress or knockdown a gene, you might need different plasmids. Harvard’s DRSC/TRiP Functional...gene to select for transformed bacteria, but you might be wondering, “How do I know which of my flies have...integration will have eyes that are anywhere from light yellow to red. The variation in color depends on...strategy and the services offered by the company, you might have to map into which chromosome the transgene ...

...neuroscience novice (newb). In this post, I’d like to highlight some of the things that popped out to me as a ...novice and indicate why other neuroscience newbies might want to get into the field. Find Optogenetics Plasmids... neuroscience but are afraid you don’t have the right kind of biological expertise to get into the field...’ve developed in your current work to make new insights in neuroscience. Even if you're currently a synthetic...interested in using AAVs because they provide a highly versatile means of delivering genes to many different...

...potential for microbial contamination. Additionally, the high activity of these enzymes implies that even a minute...a protein of interest). Mesophilic microbes are highly preferable (and successful) as host systems since...coli expressing the chaperones protein resulted in higher levels of growth compared to the E. coli strain...an outreach scientist from Addgene. The talk highlighted the importance of reagent repositories and the...leveraged for harmonious progress.     We’re always delighted to engage with potential collaborators and interested...

...is an extremely useful technique that provides insight into protein:DNA interactions. ChIP works by using...exploring these interactions, and can give valuable insight into DNA expression and protein interactions, including...require crosslinking, which uses formaldehyde or UV light to create strong but reversible bonds. This will...options for this step are sonication, which uses high-frequency sound waves to break up the DNA, or enzymatic...attach the proteins to the DNA, depending on how tightly the proteins bind natively. But the amount of cross-linking...few rounds of adjustments, to get everything just right!  Fig. 2: Goldilocks is actually celebrating...

... activation and knew she was on the right track. The next insight was to assemble a synthetic transcription...bioengineering expertise? “This is the next step in high-throughput, genome-wide screening,” Konermann said...sgRNA library. Our SAM-mediated screens exhibited a high degree of consistency and validation with >80% effectively...software for analyzing the data. It should be straightforward to apply the new library for use in various...an engineered CRISPR-Cas9 complex. Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena ...