We narrowed to 982 results for: IGH@
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TypeBlog Post...need an expression strain? Protein expression from high-copy number plasmids and powerful promoters will...typically expressed in E. coli engineered to accomodate high protein loads using inducible promoter systems (...to all or most expression strains to accommodate high protein levels including: ompT: Strains harboring... (DE3) pLysE* Chloramphenicol (pLysE) pLysE has higher T7 lysozyme expression than pLysS; expression vector... (DE3)** Streptomycin and Tetracycline Contains highly active thioredoxin reductase and glutathione reductase...—the difference being that the latter provides tighter control of basal expression. What if I don't see...media such as TB or 2XYT is optimal because of the high cell-density they support; however, minimal media...
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Prime Editing: Adding Precision and Flexibility to CRISPR Editing
TypeBlog Post...canonical base editing window, base editing has higher efficiency and fewer indels than prime editing....However, the efficiency of Cas9 cleavage is relatively high while the efficiency of HDR is relatively low, meaning...varies widely — it still almost always results in higher ratios of desired edits to indel byproducts than...average 2.3- to 5.1-fold (though up to 45-fold) higher across different genomic sites compared to PE1 ...process of double nicking increases indel formation slightly. Designing the sgRNA with a spacer that only binds...asked. Hopefully this table helps you keep them straight! Innovation Purpose PE2 PE3 PE4 ...PE6 PE7 Pentamutant reverse transcriptase Higher efficiency ✓ ✓ ✓ ✓ ✓ ✓ Second... -
Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs
TypeBlog Post...institutions of higher education nonprofit entities related to or affiliated with institutions of higher education...reserved for individuals with a US Master’s degree or higher degree. These numerically limited visas are referred...theoretical and practical application of a body of highly specialized knowledge in a field of human endeavor...the cap if the applicant works at institutions of higher education, a related or affiliated nonprofit entity...interest waiver criteria however the EB-1A requires a higher standard of proving that you are one of the top...showcases (not usually applicable to scientists). High Salary –not relevant to most postdocs. Commercial...circulation) in the field EB-1Bs historically have a high rate of approvals with USCIS due to the fact that... -
What's New in CRISPR - Winter 2018
TypeBlog Post... In this quarterly blog series, we’ll highlight a few of the new CRISPR plasmids available at Addgene...recently developed a platform called SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) to detect... this further to provide multiplexed detection, higher sensitivity, and more. Read the paper in Science...developed dual-nuclease Cas9-Cas9 chimeras with higher target site activity and more predictable and precise...inhibitor, with the LOV2 photosensor from Avena sativa, light-mediated genome editing is now possible. Learn ... -
Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping
TypeBlog Post...Proximity labelling relies on the labelling of neighboring proteins within a cell with biotin. Biotin, is...avidin coated purification techniques. Due to the high affinity of biotin for avidin, proximity labelling...promiscuous biotin ligase that can efficiently label neighboring proteins The first biotin based proximity labelling...from the ligase and bind to exposed lysines on neighboring proteins. Thus to capture protein complexes, ...electron-rich amino acids such as tyrosine on neighboring proteins resulting in their biotinylation. The...The advantage of APEX is its speed, labeling neighboring proteins in minutes rather than hours like BioID...sensitivity. Sensitivity could be increased with higher expression levels but for many fusion proteins ... -
Bringing Sustainable Practices to the Lab: Innovations
TypeBlog Post...some of these plastics are high grade and can be washed and reused. This might sound risky in terms of contamination... in a high-throughput and efficient way? Grenova, for example, offers automated, benchtop, high throughput...are an accepted part of lab culture, but with the right incentives (for example, by grant agencies), we ...behavior and innovate solutions and practices to meet higher environmental standards. Find some easy action ... -
Five Questions to Ask If You're Deciding Between a Master's or PhD Program
TypeBlog Post...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... -
Plasmids 101: Walkthrough of Addgene’s Snapgene-Powered Quality Control Process
TypeBlog Post...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 ... -
Oh, The Places You Can Go: Careers in Science Communication - Product Development
TypeBlog Post...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... -
Running for Rare Disease, Running for FOP, Running for AJ
TypeBlog Post...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... -
Advice for Choosing a Research Project
TypeBlog Post... 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... -
History of CRISPR Cas - A tale of survival and evolution
TypeBlog Post... 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... -
Antibodies 101: Avoiding the Mouse on Mouse Mess in IHC
TypeBlog Post...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... -
Intro to Our Summer SciComm Series!
TypeBlog Post...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... -
Antibodies 101: Epitope Availability
TypeBlog Post...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... -
Behind-the-scenes of the Isolation of the Thermostable IgnaviCas9 From a Yellowstone Hot Spring
TypeBlog Post... 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... -
Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?
TypeBlog Post...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... -
Hot Plasmids: FRET-Based Biosensors
TypeBlog Post...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... -
Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)
TypeBlog Post... 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... -
Virtual Science Conference Coming Up? Three Addgenies Share Their Experience
TypeBlog Post...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 ...