...fulfill our mission by generating revenue through distribution of research materials. Our small shipping and...generate revenue (as Addgene does through materials distribution). In fact, healthy nonprofits should have some...their revenue. A nonprofit organization cannot distribute profits to private individuals through stock ...request plasmids (via web interface, processes, distribution, and customer service) Addgene provides unmatched...

...the first gRNA using restriction enzyme BbsI and the second gRNA using restriction enzyme BsaI. If you’... of gRNAs Golden Gate Assembly uses Type IIS restriction enzymes, which cleave outside of their recognition...These expression vectors each contain Type IIS restriction sites flanking the promoter-gRNA construct, but...oligonucleotides. These plasmids contain Type IIS restriction sites that flank the promoter-gRNA constructs...

...protein to regulate cell growth dynamics Article contributed by Klaus Wanisch  Listen to the optogenetics ...visualizing RNA production and localization Article contributed by Tyler Ford  Listen to the riboglow podcast...developing new FPs - the mCarmine story Article contributed by Caroline LaManna  Listen to the mCarmine podcast...base-editing and Cas9 cleavage activity Article contributed by Angela Kaczmarczyk  Listen to the base editing...

...The triple T’s of biology are transformations, transfections, and transductions! In this blog we will...basics but are here because you want to know the intricacies of transductions, we've got you covered there... easy. Producer cell media can be collected, centrifuged to remove any producer cells floating in it, ...long-term (AAV). If you perform a lipid-based or electrical-based transfection, most of the nucleic acid ...

...! Archon1, a voltage sensor AAV Historically, electrical activity in the brain was measured with implanted...voltage reporter allows a way to monitor neuronal electrical activity through imaging. Since it’s illuminated...particular cell types. For example, the mDlx enhancer restricts expression to GABAergic interneurons. These vectors...pAAV-hSyn-EGFP *Also coming in our new 20 ul trial size! Voltage indicators Plasmid Serotype Name...

...This post was contributed by guest blogger Jon Backstrom, a biochemist in the Vanderbilt Eye Institute... have been immobilized on a reduced glutathione matrix. The advantage of a fusion protein is the efficient...provide a more relevant kinetic analysis, but the trick is to utilize a second immobilization method. One...available. Factor Xa loves calcium, but metal-chelate matrices, not so much. Maybe a calcium-independent proteinase...

...training progresses.  In From Academic Solos to Industrial Symphonies, an article well worth reading on ... notice or choice Always working on a team/in a matrix - can be frustrating for solo flyers Other people...—science can be surprisingly more rigorous in industrial settings Tight (sometimes impossible) timelines—others... years—a short and exciting ride Complicated matrix structures are common Small integrated teams, ...

...cloning) is a DNA cloning method that does not use restriction enzymes or ligase, and requires no post-PCR procedures...depicted as a green cloud) functions both as a restriction endonuclease and as a ligase by cleaving and ...Design standard primers (no need to add unique restriction sites on the ends) and amplify your sequence ... confirm the presence of your insert by PCR, restriction digest, or sequencing. Pro tips Do not add 5...

...This post was contributed by guest bloggers Lauren Celano of Propel Careers and Rachel Casseus, Esq. ...practice in that field. H-1B work-authorization is strictly limited to employment by the sponsoring employer...of the work of those in your field. Original contributions to the field that have had a major impact. For...of original scientific or scholarly research contributions in the field Evidence of authorship of scholarly...Addgene.org Careers at Addgene Global Plasmid Distribution ...

...of proteins for degradation using AID Article contributed by Alyssa Cecchetelli Auxin Inducible Degradation... of CRISPRCas9 by engineering SpCas9 Article contributed by Shreya Vedantam CRISPR/Cas9 is widely used...channelrhodopsins for the optogenetic toolbox Article contributed by Susanna Bachle Anion-conducting channelrhodopsins...31346176. Using LADL to loop with light Article contributed by Eric Perkins Mammalian genomic DNA is not ...

...This post was contributed by Manon Eckhardt and Melanie Brewer from the QBI Coronavirus Research Group... these proteins are FDA-approved or in clinical trials or preclinical studies already. We are following...all viral genes are flanked by EcoRI and BamHI restriction sites - so cloning projects should be fun and...partnering with Addgene, who will be taking over the distribution to expand the network of scientists working ...

...This post was contributed by Deborah Sweet, Vice President of Editorial at Cell Press. Almost everyone...Communication is the issue Much of the problem can be attributed to communication, and in particular communication...(accession numbers, reagent details, author contributions, etc.) before their paper can be published. ...across life and physical science, and editorial contribution to the strategic development of Cell Press. ...

...This post was contributed by guest blogger Lydia Morrison from New England Biolabs. What is DNA assembly...standardized DNA constructs, but its use of Type IIS restriction enzymes results in scarless assembly. Finally...allowing fragments to be taken directly from restriction enzyme digests to DNA assembly. What makes it...containing regions of homology) cut from plasmids via restriction enzyme digestion, and used directly in DNA assembly...

... exploded so quickly, citing the first clinical trial with iPSCs with his collaborator Takahashi in 2014...perspective: Dr. Yamanaka talked about his clinical trial patients with advanced macular degeneration; Laurenz...presented work that is leading to promising clinical trials for ALS.   I have an uncle with Parkinson's, a ...first stem cell line that could be used in clinical trials. The quality control process alone takes nearly...

...also use electroporation, the application of an electrical charge to cells, to increase cell membrane permeability... integrated phage remains dormant until it is triggered to enter the lytic cycle.   During both of these...they observed genetic recombination between two nutritional deficient E. coli strains that resulted in a ...cells. Electroporation on the other hand uses electrical pulses to create transient pores in the cell ...

...floats freely, depicted in the figure below. Once triggered by blue light, the CRY2 and CIB1 proteins heterodimerize...specificity as full-length Cas9 (flCas9). When triggered by blue light (470nm), paCas9-1 induced indel ...response to far-red light (730 nm). This nucleotide triggers the translocation of the protein BldD to the nucleus...constructs. This proof-of-concept model opens intriguing possibilities for optogenetic manipulation of...Gene editing Manna et al., 2019   NPPOC-caged trimethoprim (TMP) Cas9 fused to destabilizing domains (Cas9...

...resources. There are three types of cognitive load: intrinsic cognitive load is the effort associated with a...even the most enthusiastic of learners.  What contributes to cognitive load?  Everything! Here are some...specialized knowledge and context I have spent years striving to obtain?” If the answer is yes, you will need... or to gloss over some aspects when needed. The trick is to communicate your key points well enough that...

... on size is ultracentrifugation. Sedimentation equilibrium analytical ultracentrifugation yields an affinity...This post was contributed by guest bloggers Joachim Goedhart and Marieke Mastop from the Section of Molecular...on the dimerization tendency in living cells. Strikingly, the OSER assay reveals that tagRFP does not ...

...palindromic repeats separated by an 8-bp long asymmetric core spacer sequence. The asymmetry in the core...described above, a genetic modification can be restricted based on experimental constraints. This has been...site have been created, which contain a unique asymmetric spacer "NNNTANNN", where "N" indicates which ...system in your research? Do you have any tips or tricks, or clever cre-dependent techniques?  Let us know...

...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 ...