...go over caps and push them into micronic tubes instead of pushing them in one by one. It seems very simplistic...Addgene would love to move forward and automate. For instance, vial labeling would be much easier with a Tube...which air brushes the information onto the vials instead of requiring us to put a label on each tube by ...

...disease and cultured cells. For example, in 2013, Jinsong Li and colleagues chose to explore CRISPR’s therapeutic...allogenic “off-the-shelf” treatment. In short, instead of using the patient’s own cells that have been...tract infections. The Cas3 enzymes target several strains of E. coli and lethally chop their genomes into...

...replicate and continue the infection cycle once inside the host. While the virus can still enter (and ...into the host’s genome. However, some adenovirus strains are known to cause respiratory diseases in humans...your centrifugation speed, to cover the centrifuge bins. These have to be added and removed in a biosafety...

...remodeling. Chromatin is a complex of DNA and histone proteins that compacts cellular DNA into chromosomes and.... 3. Kaati, Gunnar, Lars O. Bygren, and Soren Edvinsson. "Cardiovascular and diabetes mortality determined...Experiments Post Read Other Blog Posts about Fluorescent Proteins  ...

... 5) Listen to your supervisor’s viewpoint and brainstorm about mutually beneficial solutions Your supervisor...Look at the problem from their viewpoint, and brainstorm about solutions that will meet both of your needs...important issues in person or over the telephone instead of email. As you cannot see a person’s body language...

...price point, which should be carefully balanced against the limitations of the assay.      Fig....immersion technique, courtesy of CAR15.) Each droplet contains a primer and probe set for both the gene of interest...only ddPCR provides absolute quantitative data, instead of relative quantification.  Even though no one...

...AAV does not integrate into the host genome. AAV instead relies mainly on episomal expression. Episomes ... Even more creative options involve generating strains of mice that stably express Cas effectors in specific...doi.org/10.1038/s41596-022-00700-y Hanlon, K. S., Kleinstiver, B. P., Garcia, S. P., Zaborowski, M. P., Volak...

...precise modifications of DNA (replacements, small insertions, and deletions) without introducing DNA double-strand...screens such as PRIDICT/PRIDICT2.0 provide valuable insights into pegRNA editing efficiencies for specific ...Parts, L. (2023). Prediction of prime editing insertion efficiencies using sequence features and DNA repair...

...cochlea in 3D. The basic configuration of the instrument used in this reconstruction lay the foundation...cleared whole-mount specimens, such as whole mouse brains expressing green fluorescent protein (GFP) (6)....solvent cleared organs) method of imaging rodent brains and spinal cords by combining Ultramicroscopy with...

...be better suited for the job. As single domain proteins, they can be expressed in bacteria, making nanobodies...primary antibodies from different IgG subclasses. Instead, you can pre-incubate individual primary antibodies...Learn more about protein tags Read about E. coli strains for protein expression Additional Resources on...

... Cas13 enzymes do not contain the RuvC and HNH domains responsible for DNA cleavage, so they cannot directly...functional A->G change. They fused ADAR deaminase domains (ADARDD) to dPspCas13b, but observed low RNA editing...perform C-to-U edits, the authors note that it retains its adenine deaminase ability. The authors then...

...using water-oil emulsion technology. Each droplet contains the ingredients for amplification of the target...experiment. The process to titrate AAV by ddPCR begins with diluting the virus. It is important to note...containing approximately 20,000 droplets. Each droplet contains the material required for a mini amplification...

...Our Parent Teacher Association (PTA) has been instrumental in supplementing that curriculum. It’s through...little intimidating. Too simplistic and I would insult the students. Too complicated and I would quickly... Eric J. Perkins...

...concentration of a serially diluted standard is plotted against its OD560 measurements and the trendline y = 0.6561x...the minimum and maximum detection limits of the instrument being used to measure the response. The dynamic... to set yourself up for success!  Study the instruments and specific assays you will be using ahead of...

...is introduced into the cap plasmid library by inserting short DNA sequences that contain all possible ...to 2-fold higher GFP intensity compared to the strains they were derived from. They also transduced a ...A cocktail of three AAV vectors expressing RGB proteins under the control of an inducer, i.e. tet-off ...

...that couldn’t package itself due to a few missing proteins. The envelope plasmid contained a viral envelope...lentiviral vector containing an antisense sequence against the HIV-1 envelope. As of 2013, 65 patients had...Lentiviral vectors in cancer immunotherapy. Oldham RA, Berinstein EM, Medin JA. Immunotherapy. 2015;7(3):271-84...

...credential from the Grant Professional Certification Institute (GPCI). GPCI evaluates a candidate's expertise...through the Grant Professionals Certification Institute, an Approved Trainer by the Grant Professionals... Social Media Strategist through the National Institute of Social Media. Follow Jo on Twitter @JM_Grants...

...Members of the bacterial world produce an assortment toxins to claim territory or kill competing microorganisms...an efficient cloning tool. The plasmid pKSCTMF contains the mazF gene and a gene for Zeocin resistance... James Kaper used the hok/sok system to create strains that can stably express antigens in sufficient ...

...well-supported, so we could focus on our development instead of building the entire ecosystem Proven performance...throughput necessary to keep up with our sequencing instrument (hundreds of plasmids each week!), we already...process. Similar to our first attempt, we manually inspected the output with a print statement at the end of...

... have since adapted it for a host of different proteins and small molecules from a variety of sample types...antibodies and thus are not suitable for low abundance proteins. Because the method uses a single antibody, it... and a series of wash steps removes non-target proteins and molecules from the assay. The user then adds...

...love you back A successful lentiviral infection begins with your cells, both your packaging cells and ...However, viruses pseudotyped with other envelope proteins may have a limited number of targets and therefore...Finkelshtein, D., Werman, A., Novick, D., Barak, S., Rubinstein, M. “LDL receptor and its family members serve...

... each project and how useful each item is. For instance, if I don’t have draft blog post ready for publication...prevent you from making progress on a task. For instance, even if I had planned my day extensively, my ...that highlights the things you like about it. For instance, while I may have found splitting cells boring...

...his or her chosen career path,” but individual institutions can have many different names for the positions...From Panel Session 2: Defining the Postdoc: An Institutional Perspective. This lack of complete information...Require that postdoc salaries be paid on fellowships instead of grants such that PIs no longer save grant money...

...targets, you might decide to do simultaneous IF instead (Figure 1B). To label all your targets in one step...yourself. If you can’t find a suitable antibody against your target protein, consider fusing a protein ...Neumann, E., De Caestecker, M., Gutierrez, D., & Spraggins, J. (2022). Multiplex Immunofluorescence on Fresh...

... break, or is engineered not to cleave DNA and instead activates/represses gene expression by various ...minimum of 1,000 targeted cells to sequence. In this instance, a 5% off-target frequency is something to keep...-defective lentiviral vectors - which rely on insertion of DNA by the NHEJ pathway during DNA repair of...

... a summer role as a Research Scientist at the Institute of Cancer Research in Sutton, located on the edge...and plenty of jobs in science at world renowned institutes. The lab at the University of Queensland required...field don’t really get to do a lot of lab work but instead really need to be comfortable with programming....

...transdifferentiation of glucagon-producing α-cells into insulin-producing β-like-cells [37, 38]. In 1987, Davis... ES cells and iPS cells into mature pancreatic insulin-producing cells. Cell Res, 2009. 19(4): p. 429-...duodenal homeobox gene 1 induces expression of insulin genes in liver and ameliorates streptozotocin-induced...

...used GAPDH has noted issues as a reference gene (Higgins, 2000; Ref, Zhu, 2002), though there are cases ...run and the difference in CT values is plotted against the log input concentration for each dilution. ...://doi.org/10.1038/s41598-018-26707-8Suzuki T; Higgins PJ; Crawford DR. Control selection for RNA quantitation...

...a template. To do so, they used a Cas nickase, instead of dCas9. The resulting editor, BE3, nicks the ...and/or evolved to catalyze cytidine deamination instead of (or in addition to) adenosine deamination (Chen... R. T., Christie, K. A., Whittaker, M. N., & Kleinstiver, B. P. (2020). Unconstrained genome targeting...

...Bluesky: @addgene.bsky.social; or @addgene on Instagram, TikTok, Facebook, and LinkedIn. We are also exploring...and features that previously made the platform a mainstay of conversation in the scientific community. Addgene...

...of our holiday decorations, our holiday spirit remains. We would like to thank everyone who participated...creating memories with their labmates! Are you inspired by our 2017 #Deckthelab winners? Be sure to enter...

...NanoMEDIC particles Akitsu Hotta pHCMV-RabiesG Contains Rabies virus G protein; see depositing paper for...information Miguel Sena-Esteves pcDNA-SADB19L Contains rabies viral RNA polymerase; see depositing paper...

...mTurquoise II and the creation of improved fluorescent proteins through mutagenesis. Chapter 6 - The future of...Resources from the Addgene Blog Visit our Fluorescent Proteins featured topic page Read Joachim's Series on Choosing...

...CMV promoter. Envelope proteins are a key component of lentiviruses. These proteins coat the virus and help...

...The Baker Lab focuses on designing entirely new proteins, improving our understanding of how protein folding...and others. The introduction of de novo designed proteins also helped researchers understand more about ...

...Goldilocks Antibody: Guest blogger Ava Shipman explains how the Wu lab is screening over 3,000 antibodies... a drag-and-drop CRISPR editing tool for large inserts.  DAP arrays for multiplex gene editing: Guest ...

...Plasmid Kits for microbial work with fluorescent proteins, synthetic biology, gene cloning and expression...them on our search page. Use the Expression and Insert Species filters on the right-hand side of the search...

...to cut other DNA or RNA sequences. Nucleases: Proteins that bind to gRNAs and are directed by the gRNAs...prime editing and the identification of other Cas proteins continue to expand the targeting range. CRISPR...

... about Crystal’s exciting career path, to gain insight into industry research, and to get advice on how... project and scientific teams at the Novartis Institutes for BioMedical Research to develop novel biologics...

...tradition in which we choose our teams and start brainstorming months in advance. Our teams comb the news (...Friday. So if you’re interested in finding out who wins this year’s Addgene costume contest, like us on ...

...and metabolism. Melina has worked at Genetics Institute in Cambridge, MA and Sequana Therapeutics in San...the Addgene Blog Addgene's a nonprofit? Get an Inside Look at Addgene's NGS Quality Control Addgene's...

...academic institutions can afford to hire. It will be difficult for some of these institutions to manage...

... share their AAV validation data. The Data Hub contains a curated collection of community submissions ...) can save you significant time and money. For instance, this report shows that anterograde, monosynaptic...

... about how fluorophores other than fluorescent proteins can be incredibly useful in FRET experiments. ... our posts if you have any questions or useful insights. If you're interested in blogging for us, fill...

...probably bring in a mug or reusable water bottle instead of buying yourself a big stack of plastic cups....could be: Setting up a recycling program at your institute. Set your -80 °C freezer to -70 °C and teach other...

...submission is used to create a data report, which contains experimental data showing how the antibody performed...immunocytochemistry, ELISA — but in all cases, it contains a detailed description of the experimental conditions...

...Sciences program at Harvard University and pick their brains on how they've managed to keep themselves happy...year dip (see graph). Niroshi and Ben give their insights as to what causes this dip. 11:11 - 15:37 : How...

...loads of inspiration for the current and the next generation of microbiologists. If you got inspired to produce...

... technologies were without controversy, and it remains to be seen how popular any one of them will become... though a little confusing to the uninitiated, contains much of the information needed to understand how...

...sessions led by scientists, which is hosted on our Instagram channel every Sunday. What's planned for the upcoming...participate in our one-hour Live Q&A session on Instagram every Sunday. Undergraduate and graduate students...