...off-target activity, which may be undesirable in many applications. At the time of our first blog post on choosing...CRISPR-Cas9 to increase its specificity." Nature communications 9.1 (2018): 3048. PubMed PMID: 30082838. PubMed...CRISPR-Cas12b for human genome editing." Nature communications 10.1 (2019): 212. PubMed PMID: 30670702. PubMed...

...cDNA, which  was used as a template for PCR amplification with a degenerate primer set to amplify IgG ...  While screening by IF-ICC worked well for identification of the correct clones, it was determined that...through functional assays. Following functional verification, the R-mAb expression plasmids were DNA sequenced...

...powered plasmid maps far easier to navigate. These practical and effective changes will give you the ability...promoters, origins, tags, and certain ORFs. Their sophisticated detection algorithm easily and precisely identifies...feature names, locations, sizes, colors used to indicate them on the map, direction (if relevant), and ...

... example, I will often see a whole bunch of notifications “a postdoc from this Uni viewed your profile...examples of concrete accomplishments in both the technical and “plus skills” areas. Include a professional...look straight at the camera. Pro tip: Turn off notifications to your network of profile changes. If you are...

...Not everyone in the group was able to make it physically to instruct the workshop at the conference, but...mini-grant, which will fund a project to support a dedicated website to host all the materials in one place...for me was to collaborate with this incredibly dedicated team. Many thanks to Tyler Ford, Caroline LaManna...

... the flick of a light switch. Neurons are not typically persuaded to fire when light is shined on them...directed evolution, because channelrhodopsins are typically evaluated with low-throughput methods such as ... midbrain, fiber-optic cannulas were still stereotaxically implanted into the mice’s brains so that light...

...efficient. Here, I’ll discuss transformation, DNA purification, and filling vials with agar. I’ll also provide...greater than 95% success rate! Automating DNA purification We're developing ways to prep large quantities...bacteria through the mail due to laws concerning genetically modified organisms. Since Addgene is very proud...

... and hoping to pursue. The choice to not go to medical school was devastating. At the same time, she has...circumstance. She was working as a research and technical assistant, when her life changed––she had a daughter...to your values While going through the co-op application process, my advisor encouraged me to keep my ...

...s outer membrane and allows the components to physically enter the cells so they can go to work. ...Allen Cell Explorer, where you can also find the certificate of analysis provided with each cell line. The...

...using either loxP or lox511 sites, leaving two identical sites on one end of the DNA fragment (Figure 1...recombination event then excises the DNA between the identical loxP or lox511 sites (Figure 1, bottom). Since...presence of Cre recombinase. This plasmid now specifically drives expression of mCherry instead of BFP....

...however, your cost per cloning reaction drops dramatically, as one tube of enzyme should be enough for ...there is no need for restriction digests, gel purifications, or ligations. The general Gateway cloning protocol...Chaokun, et al. "FastCloning: a highly simplified, purification-free, sequence-and ligation-independent PCR ...

...results. Most cell culture media contain the pH indicator phenol red (pH range 6.0 (yellow) - 8.0 (red))...Anything between orange and light orange usually indicates that the cells have grown enough, are making their...analysis of bisphenol A-induced perturbation in biological processes in ER-negative HEK293 cells." PLoS ...

...:  my undergrads were different people! More specifically, they had different learning styles. One was...delegating well, but it does get easier over time! Communicate often and clearly As a manager you need to clearly...it is the best way to avoid mistakes due to miscommunication and to detect trouble. I’m not comfortable...

...instances. If your poster material is nearing publication, consider uploading your work to a preprint server...time to meet. Some conferences may also have a dedicated app with a social media component, but if it doesn...other posts on scientific sharing, scientific communication, and scientific networking Learn how to set ...

...directions. Drosophila are very easy to manipulate genetically and have limited genetic redundancy (meaning,...reference of various fly phenotypes, check out the Classical Genetics Simulator website. One interesting thing... from mutations in the gene, and whether any publications have mentioned the gene. The Bloomington Drosophila...

... will have numbers by gated populations; this indicates the percentage of total events contained in the... gated population. Heat maps are often used to indicate the relative density of cells. You can see both...presented. A horizontal line across the peak will indicate the cut-off value for a positive signal (Figure...

... to synthesize a large volume of information: basically, a scientist’s tool kit. I wrote a review article... was cheating. Eventually I learned that it’s practical because there simply isn’t enough time to read...used both. Curate and present some useful data Typically lists and pictures are the most useful parts of...

...carry different antibiotic resistance markers (indicated here by plasmid color), allowing you to easily...is simple -  no restriction, ligation or gel purification steps are required! Multiple fragment cloning...the attB, P, L and R sites that recombine very specifically and directionally: attB1 sites react only with...

... use is pX333 from the Ventura lab. pX333, a modification of pX330, contains humanized wtCas9 and two ...thus marking cells that have undergone genomic modification. In this system, Cas9 must be supplied on a ... combinatorial mammalian genetics.” Journal of Clinical Investigation 125(4) (2015):1603-19. PubMed PMID...

... suggestions in the comments.  X has moved significantly away from the functionality and features that...this reason, our timeline has accelerated. We significantly reduced posting on November 20, 2023, and as...

... showed that a laughter program significantly decreased immunological markers of stress in adolescents...

...Finally, our team is rounded out with our science communication specialist (you'll know me by my Tweeting — ...leading a professional development workshop. Specifically, Jo, Brook, and Angela are co-leading "Making...

...software engineers, surprisingly, who have deep technical knowledge but little biology experience.) The ...synthetic biology, DNA assembly, and various applications. I think these papers are good starting points...

...open platform for scientists using AAV to share practical information about their experiences. It will contain...important details that you won’t find in the final publication and we have already collected over 100 entries...

...Rob Hurt, whose expertise and suggestions were critical to the development and writing of two blog posts...Modular Cloning and Plasmids 101: Modular Cloning Applications and Kits We are very grateful for each and...

...Sapozhnikov tells us how to use dCas9 (with no modifications!) as a CRISPR/dCas9-based tool for DNA demethylation...demethylation. AAV Q&A with Tim Miles: While not technically a research story, I couldn't resist adding in...

...Videos page! Videos are a powerful tool for communicating scientific knowledge. On this new page, we have...  Our first video, featuring Outreach Scientist Jessica Welch, can be found below: How-to videos Our...

...Addgene will be at the American Society for Microbiology Microbe meeting here in Boston next week - be... Jessica Welch...

... specificities, from target-specific ones to application-specific ones to anything goes. We've shared ...repositories and search engines Website Targets Application Purpose Notes Antibodypedia Any Any...

...lighting/HVAC requires a significant energy expenditure here at Addgene. These are critical systems that cannot...

...produce either a gRNA, a nuclease, or both. Applications of CRISPR: Cut: CRISPR can be used to cut the...all sequences. However, prime editing and the identification of other Cas proteins continue to expand the...

...plasmids! Each chose their favorite plasmid and replicated it. Can you guess each plasmid? Our lab paid...creative hijinks last year. They showed us their dedication to great customer support by dressing up as our...

...skipping the preparation steps of production, purification, and titering. Our newest viral vector service...E6 regulatory element, which targets it to VIP cortical interneurons. Take a look to see if these new ...

...serotype, species, or promoter) can save you significant time and money. For instance, this report shows...tropism* (min. 300 dpi, PNG format) At least two replicates of your experiment* Functional data, if the report...

...awarded plasmids from Addgene as a successful applicant to the Addgene, Seeding Labs Plasmid Grant. For...foods. 44:15 - 46:34 : Kwabena's goal to build biological science opportunities in Ghana. 46:35 - end :...

... any tags or fusions that are on your insert. Typically you would list any tag or fusion protein in the...considerations  If your insert contains a mutation or modification, this should be included in the plasmid name...

...plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools more ...monomeric Cas9’s. Read the paper in Nature Communications Find the Cas9-Cas9 chimera plasmids A modular...

...loves to visit researchers, so Outreach Scientist Jessica Welch showed some highlights of his travels from... Jessica Welch...

...'s Addgene ID # has been referenced in other publications, you'll find a link to the list of citing articles...cell-surface programmable DNA-binding domains" publication.  If you want to see this new feature in action...

...demonstrated in a report published in The Journal of Biological Engineering in March is fused to a translational...says the new tool might even find therapeutic application in addition to their use in basic biology, for...