...interference (CRISPRi) has been limited to lab-adapted bacteria because it has been challenging to introduce...the bacterial Tn7 transposon system. This system has two parts: a plasmid expressing the Tn7 transposition...for CRISPRi transfer Mobile-CRISPRi offers two advantages over existing bacterial CRISPRi methods. First...

...learn about antibodies? Our new antibodies animation has all the information! Link in bio. #STEM #antibodies...our videos will also be shared on our @addgene Instagram account, or you can watch the full animation on...

...developed for use in mammalian cells, this system has also been implemented in a variety of other organisms...Drosophila cells. Work by Wei et al. from Kang Shen’s lab has also shown that the Q system is also functional in...elegans. Subsequent work by the Luo and Potter labs has refined the Q system and introduced the second generation...multiple genes in synthetic systems. Recent work has even used all 3 in a single modular vector for ease...GAL4 gene downstream of a tissue- or developmental stage-specific promoter and design a UAS reporter construct...

...months, COVID-19 has had a tremendous impact on our world -- as of June 4, 2020, COVID-19 has caused an estimated...contains requests from 5/22/14 to 5/19/20). Each paper has between 1–32 plasmids and a total of 2–2,590 requests... 5: Inferred effects. Values are the inferred percentage of normal activity (the average rate of unique...

... its introduction in 2015 by the Boyden Lab, ExM has been steadily growing in popularity with an abundance...target concentration during expansion. One group has found that using Fab fragment secondaries, rather...experimental approaches. Since it arrived on the scene, ExM has helped researchers gain a deeper understanding of...expansion.  In some cases, labels are added after this stage.  Step 4: Expand! (Typically by just adding water...

...plasmid we have at Addgene. Each sample at Addgene has four layers of location which can all be extrapolated...hold up to 96 tubes (see image below). Each plate has numbers and letters to give coordinates, from A1 ...replicate sample. The transformation After the DNA has been received and stored, it is put into E. coli ...and prepare fresh media with antibiotics at each stage. The barcodes and LIMS help us track each sample...

...include early stage drug discovery and pre-clinical development for First-In-Man studies. Kurt has been running...Avengers This year Addgene, where my wife Lianna has made her career as Senior Director of Biology, is...

...product packaging) for free! It has to be from their company and it has to be packaging (no coated tissue...box, flip the flaps of the box so the pre-paid postage stamp is on the outside of the container, and drop...

... (SB) transposon system is one such system which has been designed to perform gene transfer in vertebrates...defective transposase found in fish. This transposase has been discovered in the genomes of many different ... both of these cases, transposons combine the advantages of viruses and naked DNA while eliminating some...

... long is the average PhD in this department? How has the department worked with students to maintain these... for everyone involved in the process. COVID-19 has changed many aspects of academia, and the PhD application...traveling for campus visits. This is definitely an advantage of surviving in a virtual world over the course...and the in-person portion got canceled, taking advantage of virtual meetings can still be a great opportunity...exception. However, using the virtual world to your advantage can strengthen your application and help you decide...

...trend in funding and space in academic research labs has more students looking for other options. While graduate... skills for alternative careers. Science Careers has developed a web-based, career-planning platform called...your project may not advance beyond its initial stages. Help you build a network outside of the academic...

... a single step. Scorpion chemistry in particular has been reported to work well in the Lightcycler while...for all amplicons; if your amplicon is AT-rich or has a secondary structure, you may see a double peak ...being a common choice - the frequently used GAPDH has noted issues as a reference gene (Higgins, 2000; ...assumptions that match your data set, that is.) Yuan, 2006 has a good overview of different statistical analysis...reaction or on a rapid cycle machine. Melt Curves An advantage of Sybr chemistry is the ability to run a melt...prevent your reaction from reaching the plateau stage after exponential amplification, making it difficult...

...extraordinary collection of talent, of human knowledge, that has ever been gathered at the White House - with the ... not an excuse. Every psych test I’ve ever taken has declared me an introvert, and I identify as one.  ... on a leaderless team. Sometimes the leader just has to be the person to say “OK, guys, we need to have...anything out the discussion themselves, then one has to question whether that person should be on the ...the first place? Let’s think about some of the advantages of team-based work: Nobody knows everything anymore...

... Lab technician / Research assistant Typically has a Master's or Bachelor's degree Generally a short-term...talking to people, though, try to be conscious of who has your best interests at heart. A great piece of advice...vulnerable about my struggles and what I don’t know yet has helped me cultivate stronger relationships with my...academia or industry, so feel free to explore! Addgene has an entire page dedicated to different career options...position taken to transition to the next career stage (ex: grad school, med school, industry) Other...

...your data to Zooniverse. The Natural History Museum has downloadable resources to enable you to set up your... doors Getting involved in science communication has been the most rewarding step I have taken in my career...: Sign up for a Twitter account. Check out the hashtag #scicomm and start following science communication...local pub at a Pint of Science event. Or get on stage for a Science Slam – a sort of competitive TED talk...

...to host proteins. After the antibody of interest has been cloned into an expression plasmid, the plasmid...through open plasmid sharing, the CRISPR community has created hundreds of useful variants of Cas9. By providing...Antibodies Recombinant antibodies offer numerous advantages over polyclonal antibodies and traditional hybridomas...AK, Anderson S, Bladen CL, Jones B, Aldred SF, Bestagno M, Burrone O, Maynard J, Ferrara F, Trimmer JS...

...invention in the late 1990s, Gateway cloning technology has become very popular as a rapid and highly efficient...attL-containing entry vector.   Pro Tip: Addgene also has ready-made entry clones available for many popular...more about the Gateway cloning method and its advantages. An introduction to Gateway technology The Gateway...and verify that your construct is functional. Advantages of the Gateway cloning method Compatibility and...JY, Chin CF (2015) Gateway Cloning Technology: Advantages and Drawbacks. Clon Transgen 4:138. doi:10.4172...

...TOPO-TA) cloning; however, the TOPO cloning technique has also be adapted for blunt end cloning. So how does...PCR products. Keep in mind that Taq polymerase has an error rate of about 1 in 3,500 bases. Typically...Resources on the Addgene Blog Perform Site Directed Mutagenesis by PCR Use REPLACR Mutatgenesis to Easily Generate...

...requiring a double-stranded break. The Xiao Wang Lab has recently deposited a set of plasmids that helps identify...in human pluripotent stem cells, a cell type that has been difficult to modify in the past.   Figure...Cas13Y, two compact Cas13 proteins found in the metagenome datasets from hypersaline samples, were engineered...

...depositing lab. If your vector is large (>10 kb) or has a low copy number, try following protocols for low...easiest and clearest ways to determine if your sample has recombined. Choose a restriction enzyme that will...contains a mixture of the full plasmid and a smaller percentage of the recombined vector backbone.   Ready...