We narrowed to 738 results for: TRI
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TypeBlog PostPublishedJune 25, 2019, 12:54 p.m....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 ...
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Optogenetics + CRISPR, Using Light to Control Genome Editing
TypeBlog PostPublishedSept. 3, 2020, 12:15 p.m....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... -
Choosing the B(right)est Fluorescent Protein: Aggregation Tendency
TypeBlog PostPublishedJune 15, 2017, 2:30 p.m.... 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 ... -
Summer SciComm Series: Cognitive Load
TypeBlog PostPublishedJune 16, 2022, 1:15 p.m....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... -
Plasmids 101: Cre-lox
TypeBlog PostPublishedJan. 13, 2015, 3:47 p.m....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... -
A Look at Addgene's QC Process
TypeBlog PostPublishedMay 6, 2025, 1:15 p.m....prioritizes the sequence quality of the plasmids we distribute to scientists worldwide. Depositing scientists...sequence discrepancies can be caught prior to distribution. Catching these errors prevents future experimental...can pass through QC and be made available for distribution. In these cases, the QC team leaves a note on...written by Alyssa Shepard, with significant contributions from Alyssa Cecchetelli, Cary Valley, and Holly... -
Quickest Way to Deposit Plasmids: The Deposit Spreadsheet
TypeBlog PostPublishedMarch 8, 2017, 3:15 p.m....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... -
Adenoviral Vector Production and Troubleshooting
TypeBlog PostPublishedJune 14, 2018, 11:44 a.m..... The main challenge with using AdV is that it triggers a strong immune response after in vivo administration...personalized medicine (2017). 1st Gen AdVs are stripped of regulatory genes E1 and E3. Without these genes...plasmid stock should be analyzed by endonuclease restriction digest or fully sequenced to confirm its integrity...chloride (CsCl) density gradients combined with ultracentrifugation to separate rAdV from other cellular debris... -
High Complexity Golden Gate Assembly with NEB’s DAD
TypeBlog PostPublishedAug. 1, 2023, 1:15 p.m....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 ... -
Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them
TypeBlog PostPublishedMay 4, 2021, 1:15 p.m....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... -
Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors
TypeBlog PostPublishedAug. 20, 2019, 12:31 p.m....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... -
Adapting Toehold Switches to Detect Zika Virus
TypeBlog PostPublishedAug. 30, 2016, 2:30 p.m....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... -
Hot Plasmids: Winter 2025
TypeBlog PostPublishedMarch 11, 2025, 1:15 p.m.... 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... -
Pooled Library Amplifications
TypeBlog PostPublishedMay 30, 2024, 1:30 p.m....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 ... -
10 Basic tips for mammalian cell culture
TypeBlog PostPublishedJuly 12, 2018, 1:09 p.m....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... -
No Llamas Required - Synthetic Nanobodies Against Membrane Proteins
TypeBlog PostPublishedJune 18, 2020, 1:15 p.m....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... -
Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection
TypeBlog PostPublishedDec. 12, 2013, 3:35 p.m.... 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 ... -
Great Results Start with Great Standard Curves
TypeBlog PostPublishedAug. 15, 2024, 1:15 p.m....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. ... -
AAVs CREATed for Gene Delivery to the CNS and PNS
TypeBlog PostPublishedSept. 28, 2017, 2:01 p.m....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... -
The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data
TypeBlog PostPublishedJuly 16, 2020, 1:15 p.m....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...