...may include reduced saturation density, decreased rates of growth, or clumping. To ensure the best possible... culture-based experiment, labs should routinely test for mycoplasma contamination. There are several ...with freely available positive controls. Routinely test your cell line for mycoplasma and discard it if ...routinely produce lentivirus may want to consider testing a variety of cation and serum sources to ensure...

... (pLemo) Rhamnose-tunable T7 RNAP expression alleviates inclusion body formation. Expression vector cannot... recA+, KmR Expression of toxic proteins  * Denotes the presence of an additional plasmid-- make sure...setting up a small-scale expression experiment to test variables such as temperature, time, and media conditions...temperature or moving it to a bench-top shaker 10-20 minutes before adding the inducer. Growth Media: Changing...

...tightly regulated. Common promoters for eukaryotes and prokaryotes We have included two reference tables...bacteria this is done by a single enzyme; however, eukaryotes have muliple polymerases which are each responsible...expression mRNA Yeast promoter containing Gal4 binding sites, commonly used in Drosophila Specific Requires the...

..., followed by an intramolecular reaction that generates the chromophore (Tsien, 1998). Importantly, this...absorb light at 488 nm. The ratio between the two states is reflected by the pKa, which is around 6.0 for...relatively small, adding GFP may alter diffusion rates. Smaller genetically encoded probes are based on...Adjobo-Hermans, 2011). Structural information facilitates the choice of the proper site, although it still...

...approach is most well suited, since it routinely generates knockin efficiencies of 1-40% with G418-selected... AAV to deliver their repair template to mouse zygotes and thereby correct a mutation that causes Hemophilia...technique to efficiently deliver large repair templates for applications that require large gene fusions...Porteus. "CRISPR-mediated integration of large gene cassettes using AAV donor vectors." Cell reports 20.3 (2017...

...different splice site donor and acceptor sequences were tested and with the optimal sequences, the group achieved...homologous recombination approach delivers large donor templates using two sequential homology directed repairs...2017) CRISPR-Mediated Integration of Large Gene Cassettes Using AAV Donor Vectors. Cell Reports 20:750–756...PA (2002) An mRNA Surveillance Mechanism That Eliminates Transcripts Lacking Termination Codons. Science...

... Fig 1: Quote from Michael Ende, courtesy of morefamousquotes.com A good listener absorbs and remembers...college, when I discovered that many of my old suitemates stopped talking to each other if I was not there... diagram of the Thomas-Kilmann Conflict Model, courtesy of https://managementweekly.org/thomas-kilmann-conflict-resolution-model...

...molecules, knowing that where you see fluorescence indicates your target’s location. Some other terms you might... The biggest difference between IF and ICC/IHC relates to the type of secondary antibody you use, so as...corresponding protocols. Other researchers may also have tested an antibody in IF and will share their findings...of interest. Thankfully, there are a number of websites compiling antibody data that can help you compare...

...-target sites with up to 7 mismatches to the gRNA target sequence. Kaminski et al. next tested their procedure...targeting of the enhancer region near the NF-KB binding sites increased activation of HIV promoter-luciferase ...expression of Cas9/gRNAs, which could result in higher rates of off-target cleavage. In both cases, translating...

...not require the presence of specific restriction sites (or lack thereof) nor a serious time commitment....of fragments, but uses site specific restriction sites (Type IIS restriction endonucleases) to cleave DNA...that the vectors and DNA fragments contain these sites at the correct location and NOT in the middle of...flanked by orthogonal versions of the attB and attP sites recognized by the integrase. This method requires...

...Vectors via Different Routes. Diabetes, 55(4), 875–884. https://doi.org/10.2337/diabetes.55.04.06.db05-0927...delivery by directed evolution of underexplored sites on the AAV capsid. Molecular Therapy. Methods & ...

...(Wenning) was fascinated by CRISPR and rushed to test it. I still remember the day, March 16, 2014, that... it has been repurposed for genome editing in eukaryotes, with the most widely used CRISPR genome editing...specifies the target, the Cas9 endonuclease that creates the DNA double-strand break (DSB) at the target...mouse zygote (Qin et al., 2015).  When inside the zygotes, the gRNA will seek out its target among the 3 ...

...country of origin with an international mentor creates a closer and more approachable relationship for...evidence of this, you needn’t look any further than testimonials from the students who participated in the program... of Oviedo, the European University Miguel de Cervantes, the University of Navarra, and the University...macular degeneration. She is also a blogger who writes posts for the blogs of MasScience, ECUSA, IMFAHE...

...the surface of extracellular pathogens such as parasites or microbes, or to proteins expressed on the surface...cascades that clear these infections. Anything that generates an antibody response in the immune system is referred...at different times of the immune response and initiates different immune cascades. In the research setting...quantitative measurement of protein expression in cell lysates, whole cells, or tissue samples. Many experiments...

... announce the winners of our 2021 Deck the Lab Contest! Though 2021 had more than its fair share of challenges... challenges, the annual Deck the Lab Contest brought us smiles and joy. In fact, our Addgenies were absolutely...festive makeover today✂️❄️🦠thanks to the awesome templates by @CVRHutchinson @CVRinfo. Check them out: https...

... start looking to find one. Luckily, there are websites that can help: Antibody data repositories, which... Website Targets Application Purpose Notes Antibodypedia Any Any Data repository ...Molecules group Addgene Data Hub Any (see notes) Any Data repository Only accepts data for...

...through 2009, though I suspect that our costume contests were indeed occurring earlier but that the photo...protect the innocent revelers. Our Halloween costume contests have become an annual tradition in which we choose...finding out who wins this year’s Addgene costume contest, like us on Facebook and checkout our FB page on...

...our in-stock service: ddPCR, purity testing, and sterility testing, plus viral genomic sequencing to confirm...AAVrg and 7 × 1012 GC/mL for AAV2. There are no quotes or special processes involved; simply request your...

...only March!)  In our repository, you'll find the latest iteration of GCaMP, GCaMP8, available in new serotypes...repository  Name Serotype Category Depositor  Notes AAV-CAG-jGCaMP8s-WPRE AAV1 Biosensor Looger New...forgot to use the link below to keep up with the latest in Addgene's viral vectors services and resources...

...you need access to raw data, the target sites, off-target sites, scores and the statistics that go into...genome contains only 0.66 million targetable NGG PAM sites whereas the human genome has about 300 million. ...newer tools may still prove somewhat useful for the latest NTT targeting Cas protein, Cpf1. Separately, on-target...