...the Doyon lab has also provided a detailed protocol at Protocol Exchange. Plasmid maps can be found at Addgene...cell types directly linked to disease, including motor neurons (-3mnx1/Hb9), pan neuronal (elavl3/HuC),...mScarlet-H with M164H (2-fold improvement in photostability compared to mScarlet, fluorescent lifetime ...Listen to our mScarlet Podcast Segment    Photocleavable protein PhoCl expands optogenetics toolbox ...shift variant, which could be compatible with two-photon microscopy using Ti-Sapphire lasers. pBAD-RDSmCherry1...

...on how to apply the new system, see Yamamoto and Sakuma’s protocol. Reference: Multiplex genome engineering...filled that gap. As Sakuma and colleague Takashi Yamamoto describe in their recent study, the construction...system. Sakuma T, Nishikawa A, Kume S, Chayama K, Yamamoto T. Sci Rep. 2014 Jun 23;4:5400. doi: 10.1038/srep05400...

...fight against cancer in its many forms. The dichotomous interactions between tumors and the immune system...recognized by effector cells of the immune system - cytotoxic T lymphocytes (CTLs), natural killer (NK) cells...programmed cell death ligand (PD-L1) that inhibits the cytotoxic activity of CTLs and NK cells. These cells would...immunotherapy. Using activated DCs ex vivo to generate cytotoxic effector T cells, and targeting specific DC subsets... targets like CD19 on tumor cells and exhibit cytotoxic activity. CAR-T cells have consistently shown ...

...Although ChIP-seq is a relatively simple and robust protocol it does have limitations. The enzyme-based CUT... CUT&RUN method Enter CUT&RUN, an enzyme-based protocol for identifying DNA binding patterns that overcomes... outlining improvements to the initial CUT&RUN protocol, the most significant being the usage of a strong...Additional Resources on Addgene.org Find simple lab protocols Browse CRISPR plasmids for isolating a genomic...

...advantage of bacterial characteristics such as prototrophy, the ability to synthetize all compounds needed...synthesize a compound needed to grow. Auxotrophy and prototrophy come into play during cloning if you’ve designed...example, Markus Ralser’s lab created a yeast prototrophy kit containing 23 plasmids to complement auxotrophy...molecular biology reference Watch molecular biology protocol videos ...

... purification or if you use a DIY purification protocol, you might have noticed that there are many options...options to elute your DNA prep. You might see protocols that recommend eluting in water, Tris-EDTA (TE...agents that can be introduced in the purification protocol to remove endonucleases activity. For these reasons...Check out the Molecular Biology Reference Find protocols on plasmid cloning Find plasmids for your research...

...start tagging? We are here to help with vectors, protocols, antibodies, and more!   Resources and References...., & Wingfield, P. T. (Eds.). (2001). Current Protocols in Protein Science. John Wiley & Sons, Inc. https...In P. E. Vaillancourt, E. coli Gene Expression Protocols (Vol. 205, pp. 99–118). Humana Press. https://...antibodies Addgene’s antibody guide Antibody protocols  Resources on the Addgene blog Plasmids 101:...

...Science Resources on Addgene.org Find Protocols and Protocol Videos Check out LeGO plasmids from the... very human side to scientific research. Find Protocols Lab Tips to Keep Your Experiments Running Smoothly...this world where we are often trained to follow protocols, guidelines, and standards. Bricking science reminded...

...introductory blog post, then check out this RNP protocol on Protocols.io. The PAM Requirement and Expanding CRISPR...Channabasavaiah B. Gurumurthy, and Masato Ohtsuka. Nat Protoc. 13(1) (2018):195-215. PubMed PMID: 29266098 Creating...models with CRISPR has been challenging, with protocols using dsDNA templates achieving ~1-10% efficiency...

...Plasmid Cloning Protocols Catch Up On Your Basic Molecular Biology Techniques Watch Our Protocol Videos ...interest with Taq polymerase using your favorite PCR protocol.  2. Set Up TOPO Cloning Reaction: Mix together...Competent Cells: You can use your standard lab protocol for this; however, you should reduce your incubation...

... fly library web page offers more details and protocols, and the Liu lab’s OXfCRISPR page provides many..., pCAG-Golden-Gate-Esp3I-Destination Takashi Yamamoto pcDNA-TAL-NC2, pCAGGS-TAL-NC2 Charles Gersbach...than common lab yeast strains since they are prototrophic, usually have low levels of homologous recombination...markers, allowing for use with both laboratory and prototrophic strains. The vectors were based on the original...PMID: 27579989 DECEMBER 2016 Restoring yeast prototrophy with plasmids Though auxotrophic markers are ...lab has created a unique tool with their Yeast Prototrophy Kit, a set of plasmids designed to complement... and besides being used for restoring strain prototrophy, can also aid in designing self-establishing ...

...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...

...organized allowing you to upload anything from protocols to pictures of gels or even sketches to your notebook...but the NEB Tools app comes in handy when I use protocols that require restriction enzymes or PCR. The app...Bioinformatic Analyses Use Apps to Share Your Lab Protocols Check Out How Addgene Develops Lab Management ...Management Software Resources on Addgene.org Find Protoctols for Your Research Catch Up On Your Molecular Biology...

...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...

...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...

...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...