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...Cells were cultured following a differentiation protocol before performing single-cell RNA sequencing. ... refer to the Zhang lab’s previously published protocol and its relevant modifications on the pooled library...looking for the best astrocyte differentiation protocol! Ready to try it out for yourself? Check out ...knockout and transcriptional activation screening. Nat Protoc. 2017 (4):828-863. doi: 10.1038/nprot.2017.016....

...standard lentivirus production protocol here and you can find more tips and protocols for working with lentivirus...researcher can complete an entire lentiviral production protocol only to find that the low titer of her prep makes...lentivirus on our blog and protocol pages. We’re excited to provide this new service for researchers wanting...

...explore how engineering fields rely on rapid prototyping to develop and test hypotheses. Students then...then use cellular extracts as a means to rapidly prototype DNA circuits and logic gates. This BioBuilder-...projects at any stage of completion, from pitch to prototype. Last year’s BioBuilderClub teams’ biological ...the Importance of Assembly Standards Synthetic Photobiology: Optogentics for E. coli Browse All Synthetic...

... efficiency by which absorbed photons are converted into emitted photons. These are indicated by the extinction...this series of posts are (i) brightness, (ii) photostability and (iii) aggregation tendency. Brightness...Without further information about properties like photostability or aggregation tendency, our choice would be...

...helping us establish consistent, high-quality protocols for lentiviral production. Bryan Roth’s lab from...technical service (details about virus production protocols and quality control can be found on Addgene’s ...and Office Teams develop shipping systems and protocols for viral vectors to reach scientists around the...Beginners Resources at Addgene.org Find Virus Protocols Visit Our Viral Vector Pages Find Virus for Your...

...that are currently available along with detailed protocols, go to www.addgene.org/crispr/tagging. Addgene...number of groups were excited and asked for the protocol. Since the Addgene page went up last month, over... to their own work? Mendenhall: We’ve made the protocol available here. There were a few bumps we went... in the lab already should be able to run this protocol as well. We would also like to put some social...

...indel generation for two PTG constructs in rice protoplasts. Compared to matching traditional gRNA-expressing...3 and 31 times higher, respectively. In rice protoplasts, Xie et. al observed slightly higher mutation...mutation rates than they’d previously reported in protoplasts (15% and 9% of target sites.) How many loci can...gRNAs targeting 4 MAPK genes, again using rice protoplasts. Each pair of gRNAs targeted genomic sites separated...

...fluorescence microscopy, flow cytometry, or spectrophotometry. The range of fluorophores available for antibody... et al., 2007; Stoeckius, et al., 2017). Metal isotopes are useful for mass cytometry (Bendall, et al....Modifications in Cells and Tissues in Situ. Curr Protoc Cell Biol 89:e115. https://doi.org/10.1002/cpcb...Detecting antigens by quantitative immuno-PCR. Nat Protoc 2:1918–1930. https://doi.org/10.1038/nprot.2007.267...

...Technique Find Plasmid Cloning Protocols Check Out All of Our Protocols Additional Web Resources OpenWetWare's...Addgene Blog for a variety of useful resources and protocols! We invite feedback from all members of the research...killer protein: ccdB gene mutants coding for non-cytotoxic proteins which retain their regulatory functions...

...-radiative transfer of energy (i.e. no emitted photons) from the excited donor fluorophore to the acceptor...Donor fluorophore excitation by absorption of a photon 2. Energy transfer from the excited donor directly... acceptor fluorophore--think of it as a virtual photon 3. Relaxation of the acceptor fluorophore back ...back to its ground state by emission of a photon with wavelength specific to the acceptor In order for FRET...

...instance, we use a mutant known as CX51 that exhibits motor defects above 25 degrees. Computer scientists and...CRISPR/cas9 to build the mutants, analyse their motor behaviour, and ultimately submit our new strains...our final target decisions, we intend to focus on motor neuron knockouts so that we can perturb factors ...our members took an Arduino microcontroller and a motor controller and made it spin again!  We’ve also learned...

...thinking of an amazing project. You’ve written your protocols, designed your experiments, and prepared your ...purifications, or ligations. The general Gateway cloning protocol is fairly flexible and adaptable to cloning different... they will come with comprehensive and tested protocols, as well as premixed and ready-to-use reagents...101 eBook Resources on Addgene.org0 Check out protocols for plasmid cloning Find vector backbones Read...

...details on this procedure check out our PCR protocol page, protocol video and reference on how to design PCR...Resources on Addgene.org Protocol: Polymerase Chain Reaction (PCR) Protocol: How to Design a primer  ...

... targeting vectors. Addgene depositor Takashi Yamamoto’s lab has harnessed MMEJ to create a new method...previous work, Sakuma et al. describe a detailed protocol for MMEJ-mediated knock-in of a GFP-Puro cassette...much shorter sequence homologies. Abbreviated protocol for PITCh Step 1: Generate microhomologies in ...TALENs and CRISPR-Cas9 with the PITCh systems.” Nat Protoc. 11(1): 118-33. PMID: 26678082. Find the plasmids...

...order to study the conformational dynamics of the motor protein kinesin-1, the Twelvetrees Lab took advantage... molecules, which are much brighter and more photostable than fluorescent proteins, making them ideal ...versions. ASAP5 can be imaged using one- or two-photon microscopy to detect action potentials and subthreshold...fluctuations in awake behaving mice through two-photon microscopy at depths up to 600 µm. Figure...

...cell level as they are in vitro. Factors like photobleaching, lack of red-shifted indicators, and brightness... linked to a chromophore and functions as a photoreceptor in the eye. There are two flavors of rhodopsin...issue with these GEVIs? They perform poorly in two-photon conditions, which are necessary conditions for ...Addgene.org Addgene’s Viral Services Addgene’s Virus Protocols Addgene’s Chemogenetics Guide Resources on the...

...guidelines here. A detailed transformation protocol, as well as protocols for growing Arabidopsis are also available...more than one construct at a time. For detailed protocols, volumes, spin speeds, recipes, and Agrobacterium...your insertion in the absence of selection. For a protocol for antibiotic selection, check out Harrison et...

...enhanced propensity for movement and activity called locomotor sensitization. This enhanced movement is thought...craving (‘seeking drugs’).  What’s involved in locomotor sensitization? Previous research has shown that... previously suggested inhibitory mechanism of locomotor sensitization and might open new ways for a novel...Learn about Addgene's viral service Find viral protocols and videos Find AAVs for your research ...

...crisis. I like how they are using protocols.io to create a protocol community! Asclepias syriaca and Asclepias...solicitation requires funded labs to rapidly disseminate protocols, methods, data, and tools that result from their...Chisholm) This cyanobacterium is the smallest photosynthetic cell in existence, but most abundant cell on...

... tries to “optimize” a particular standard lab protocol. For example, someone may attempt super quick ...quick incubation times in a Western Blot protocol and then blame the antibody when no signals are obtained...so many things which can go wrong in a lengthy protocol. And customers may tell all sorts of details about...reviewing customer data sets, troubleshooting protocols, analyzing information, and solving customer problems...