...into the liquor of the cerebral ventricles (ICV, intracerebroventricular) to minimize tissue damage; however...Experimenter Interaction should be minimal and strictly controlled; optional handling animals Experiment...depending on the choice of channel protein, they can trigger neuronal excitation or inhibition upon light (or...

...that the five traditional rules didn’t need to be strictly followed in order to generate high-fidelity reactions...overhang set by uploading the set with the Type IIS restriction enzyme and thermocycler protocol. It will also...a circular or linear assembly. If you aren’t constricted to a specific number of fragments, you can also...correctly assembled phage genomes, it required a very stringent selection process and benefited from a longer ...

...data (Begley & Ellis, 2012; Baker, 2015). This contributes to an enormous amount of wasted money and time...Nguyen K, van der List D, Murray KD, Engebrecht J, Trimmer JS (2019) A toolbox of IgG subclass-switched recombinant...https://doi.org/10.1038/520295b Bradbury ARM, Trinklein ND, Thie H, Wilkinson IC, Tandon AK, Anderson ..., Bestagno M, Burrone O, Maynard J, Ferrara F, Trimmer JS, Görnemann J, Glanville J, Wolf P, Frenzel A...

...in any format. Distribution status for your plasmids: Hold for Publication or Distribute pending QC. The...Gibson cloning, ligation-independent cloning, restriction enzyme, TOPO cloning, unknown, or other. ...or licenses that could prohibit Addgene from distributing the plasmid, select “yes” from the drop-down...

...This post was contributed by guest bloggers Keith Pardee and Alexander A. Green. Zika background First...successful sensors to draw on as our assays grew more stringent, and it avoided any days of lost productivity ...when we needed additional expertise. Perhaps most striking is that there were seven institutions involved...and open approach by all involved definitely contributed to our ability to complete the project on such...

...This guest post was contributed by Sana Khan Khilji, a PhD student at the Max Planck Institute of Colloids...that UV light only kills microorganisms that it strikes directly. UV light exposure is also harmful for...way to prevent contamination. This sounds rather trivial, but often solves most problems before they occur...PubMed Central PMCID: PMC4047077. 2. Pagliara, Patrizia, et al. "Ostreopsis cf. ovata induces cytoskeletal...

... functional heterotrimers with endogenous Gβγ, enabling detection of activation triggered by endogenous... responsivity in screening ASAP3 variants. B) Electrical and optical recordings of spontaneous activity...sharing your experience with other researchers by contributing to the Addgene Antibody Data Hub.  Figure...

...and loss of plasmid diversity, so Addgene strives to distribute aliquots of the original library whenever...cheaper and easier. One library that Addgene distributes with a chemical transformation amplification ...with. In either case, it can be useful to run a restriction digest of your amplified library on a gel to ...

...antibodies against proteins embedded in membranes is tricky because they have fewer hydrophilic surfaces, require... randomized with a defined set of different trinucleotides (corresponding to different amino acids) in...exchange (FX) cloning technique which uses Type IIS restriction sites (Geertsma, 2013). Once the phagemids are...after a few washes, binding is detected using a colorimetric reaction. After the ELISA step, the narrowed-down...

... this way: "Addgene is one of the leaders in distributing plasmids amongst the academic community. On ...has partnered with the BioBricks Foundation to distribute the physical DNA samples, for when that's the...central dogma at the genome scale, it's going to be tricky to tame the rest of a cell. The BIOFAB plasmids...Who owns computer-programming languages? That's trickier. Sometimes it's the developers. But, it turns ...

...dilution series should be run in duplicate or triplicate. The closeness of the data points generated by... the standard dilution series in duplicate or triplicate. Replicates allows you to calculate the curve...within the linear portion of the graph. It does restrict the dynamic range of your standard curve, so if...and mix samples very well.  Use duplicates or triplicates to increase the accuracy of your experiment.  ...

...Vector  Percent of Neurons Transduced   Cortex Striatum Cerebellum AAV.PHP.B (parental) 49 +/- 7% 36...of a transgenic mouse. 2. Cell-type-specific restricted transgene expression in the CNS and PNS Previously...promoter or enhancer now allows for cell-type-restricted expression in both the CNS (figure 5 in Chan ...These capsids can serve as powerful cell-type-restricted gene vectors for neurons, no transgenic mouse...

...This post was contributed by Max W. Shen from MIT, Alvin Hsu Harvard University, and David R. Liu from...statistics provided by Google on 6/4/20). Many countries entered lockdown for weeks to months, pausing ...requests. We received plasmid requests from 56 countries, though the data is heavily dominated by the U.S...noise to each of the terms which make up our distribution, resulting in the following hierarchical model...

...solution. Proteins and genomic DNA are removed by centrifugation. At this point, the plasmid DNA sample can ...binding DNA, the glass particles are pelleted by centrifugation and washed before DNA is eluted with water ...was free of agarose and could be digested with restriction enzymes.  Cellulose nucleic acid purification...phosphoric acid for 3 minutes followed by a short centrifugation for a total of four times before a wash with...

...decided this protocol would be a worthy first contribution to the protocols.io community. Did I mention... protocols, including making LB agar plates, restriction digest of plasmid DNA, and bacterial transformation... was created in 1999 and allows scientists to contribute protocols or provide links to protocols on their...subscription-based software, Labguru does offer free trials and pricing options for academic labs, departments...

...made great strides to map brain regions and allocate specific functions to them. Electrical recordings...circuit by integrating neuronal signalling and triggering release of the stress hormone norepinephrine ...levels in general, or anxiety associated with psychiatric diseases such as schizophrenia or bipolar disorder...

...protein ticker tape Teaching an old antibody new tricks   New compact, efficient prime editors By Susanna....10.13.463862.    Teaching an old antibody new tricks By Ashley Waldron While IHC protocols typically...thick-section and whole-mount labeling techniques. The Trimmer lab’s anti-PSD-95 [K28/43R] recently passed their...antibodies in Addgene’s collection since we began distribution and has been verified to perform well in standard...

...single crRNA The Feng Zhang lab used Cas13a from Leptotrichia shahii (LshCas13a) to begin to characterize ...et al., 2017). They characterized Cas13a from Leptotrichia wadei (LwaCas13a), using a superfolder GFP fusion...global RNA expression, cell stress, and RNA size distribution. When comparing LwaCas13a to shRNA, they found...community. Note: Cary Valley and Mary Gearing contributed to the update of this post. References 1. Abudayyeh...

...envelope proteins from another virus to either restrict or broaden the host cell range (tropism). Pseudotyping...vesicular stomatitis virus G protein. VSV-G is a trimeric protein that binds phosphatidylserine and low-...preparation, VSV-G is stable and can withstand ultracentrifugation which helps scientists achieve a high titer..., Young JAT, Callaway EM (2007) Monosynaptic Restriction of Transsynaptic Tracing from Single, Genetically...

...and last updated Aug 12, 2021. This post was contributed by guest bloggers, Yiwen Zhang, a Ph.D. candidate... pTargetF with the ccdB gene containing BsaI restriction enzyme sites at its ends to generate pEcgRNA....that is 2.6 times higher when compared to the industrial strain. In our metabolic engineering case study...high-yielding strain which had 6 times the yield of the industrial strain. Not only is MUCICAT better, it is faster...