We narrowed to 618 results for: abo.2

...CombiGEM lentiviral vector backbone pAWp12 (Figure 2). Cloning from the pAWp28 storage vector to the pAWp12... before beginning your experiments. Figure 2: Strategy for assembling barcoded combinatorial gRNA...PMID: 25114216. PubMed Central PMCID: PMC4151723. 2. Doench, John G., et al. "Optimized sgRNA design to...How Barcodes Can Be Used in Lineage Tracing Read about CRISPR/Optogentics Tools Resources on Addgene.org...

...require application submission in-house approximately 2 weeks before the RFA due date in order for the administrative...deliverables to the aims requested in the call. Step 2: Develop a research plan Most grant applications ...proposal typically lists the deliverables, explains 1-2 potential pitfalls of the intended protocol, and offers...first. Additionally, reach out to peers and collaborators who have succeeded at grant writing to see what...

...retinal cell-type specific genes from mice. Strategy #2 (ProB): Create promoters using phylogenetically conserved...would express Cre-mCherry and GFP.   Figure 2: Synthetic promoters can be paired for targeted expression...1345-1356. https://doi.org/10.1038/s41593-019-0431-2  Neff, E.P. An AAV library for retinal cell expression...Addgene blog Read more about using AAV for retinal gene therapy Find more about precise neuronal targeting...injected into macaques and postmortem human retinas. About 25% of these AAV synthetic promoters successfully...

...our deposit spreadsheet to complete Step 1. Steps 2 and 3 are usually very easy — our tech transfer team...columns are shown in red in the spreadsheet (Figure 2) and are described in Table 1. Some cells have drop-down...Filling out and sending the deposit spreadsheet Figure 2: Example of the deposit spreadsheet showing the first...streptomycin, tetracycline, combinations of the above, or other. High or Low Copy Choose from: high...the “Purpose” field. If you ever have a question about what should go into a cell, hover over the cell ...Addgene and contact your technology transfer office about the MTA. Pro Tip! Download a new spreadsheet each...

...AAVs in the clinic Thus far, AAV vectors AAV1, AAV-2, AAV1-AAV2 hybrids, AAV-6, AAV-7, AAV-8, AAV-9 and...neutralizing antibodies against some AAV types, AAV1, 2, 3 and 5 are thought to be as high as 70% among the... 10.24 (2003): 1999-2004. PubMed PMID: 14566358.  2. Hirsch, M. L., et al. "Self-complementary AAV mediates...Vectors Featured Topic Page Learn about CRISPR-AAV vectors Learn about Addgene AAV Quality Control Process...viral infections. She is particularly passionate about HIV cure research and science communication.     ...

...Reexamining assumptions about miRNA-guided gene silencing. Nucleic Acids Research, 50(2), 617–634. https://...downstream genomic sequences (Figure 2B).   Figure 2: A comparison of an shRNA plasmid sequence and an ...module_attribute "schema_version" is_json="true" %}{% raw %}2{% endraw %}{% end_module_attribute %}{% module_attribute...References Bartel, D. P. (2004). MicroRNAs. Cell, 116(2), 281–297. https://doi.org/10.1016/s0092-8674(04)00045...

... of components, such as your CRISPR enzyme or Pol 2 promoter. The options available in each class, or ...overhang specific to the type of module it is (Pol 2 protomer, Cas protein, ect...). Whatever individual... (KO, CRISPRa, CRISPRi, editing)   Figure 2: Selecting module fragments on the Fragmid website...Destination vector (dropdown options depending on above selections) Module set (the set of components you...vectors) Target type (drop down options depending on above selections.) CRISPR enzyme (Cas9 [sp], Cas9 [other...

...neoblast cells and the gene Prohormone convertase 2 (pc2) labels the central nervous system. Evaluating... characterized in an unrelated worm (C. elegans) [2]. Delivering dsRNA matching a specific mRNA sequence...223). Link: http://abt.ucpress.edu/content/79/3/208 2. Fire, S. Xu, M.K. Montgomery, S.A. Kostas, S.E. Driver...experiments in the beginning. In most classrooms, the laboratory experience is focused on classical embryology... Biology with BioBuilder 3D Models for learning about CRISPR Resources at Addgene.org Find plasmids ...

...the system outlined in Schmid-Burgk, J.L., et al (2). In this post, I’ll cover two great applications ...molecularly evolved from mNeptune2 through 33 mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness...luciferin 24 hours later demonstrated a more than 2-fold brighter signal from Antares as compared to Fluc...PMID: 22894855. PubMed Central PMCID: PMC3501149. 2. Schmid, J.L., et al. (2016) CRISPaint allows modular...Blog Learn more about FRET Tips for Using FRET in Your Experiments Learn more about Fluorescent Protein...heme-containing proteins in whole animals, light at or above 600 nm wavelengths should be used. Researchers often... emits light at a tissue-penetrating wavelength above 600 nm. LumiFluors Schaub, F.X., et al. (3) designed...

...design primers to detect the presence of your insert; 2) set up a standard PCR reaction (primers, dNTPs, polymerase...strategies for primer design: 1) insert-specific primers, 2) backbone-specific primers, and 3) orientation-specific...negative clone (-) will not. Backbone-specific primers (2) give larger sized products for positive clones (+...SLIC cloning Learn about Gibson Assembly Additional Resources on Addgene.org Read about Choosing a Molecular...this type of primer does not provide information about the orientation of the insert. Orientation-specific...Orientation-specific primers: If you need information about insert orientation, then you might consider designing orientation-specific...insert Backbone-Specific Provides information about the size of the insert and if it's in the plasmid...

... then overlaid with the firmer 2% agarose gel solution discussed above for picking. Many thanks to our...3 plates at different densities for selection). A 2% agarose solution is then melted (and thereby sterilized...96-well assay plate. We generally freeze at least 2 ampules each of 4 clones with different levels of ...specific genes. Many years ago, people in the laboratory became frustrated with using glass cloning rings...the hood. A 48-well plate is also prepared with about 200 ul of culture medium containing serum in each...incubator, additional growth medium is added to about halfway up each well. Tips for using the agarose...

... characteristics of select AAV serotypes in Table 2 from the paper. Delivery of AAV into the brain can...routes for neuroscience see Table 1 from the paper. 2. Manipulate your cell of interest: Cell-type specific...Learn more about retrograde AAVs Resources on Addgene.org Read our AAV guide Learn more about our viral...

...digestion of the destination vector with SapI yields 2 sites- the first site accepts the sgRNA target sequence...marker, and connector to be released (Fig. 1, Step 2-3). Since donor plasmids of the same type will produce... a gRNA vector that targets the gene of interest, 2) a plasmid to specify the reading frame of the insertion...tagged gene. Most insertions can only be detected by labor-intensive processes like PCR or by evaluating visual...repair after a CRISPR/Cas9 induced break. Learn more about these constructs and the cell lines they’ve been...Additional Resources on the Addgene Blog Learn about Fluorescent Protein Timers Check out Photoactivatable...

...the mixed sample in our pipeline (see Figure 2, sample 2). When we blindly analyzed AAV-68544, we were...look for potential contaminants. The analysis is a 2-step process using Geneious software: First, the individual...viral genome sequencing (VGS). Why should you care about viral quality control? AAV vectors are produced ... let us know if you have any specific questions about viral DNA NGS you’d like us to discuss. We are also...or at [email protected] to answer your questions about our quality control process! The images in this...

...base pair to another base pair at a specific site. 2. Temporal, spatial, or conditional control of CRISPR...difficult-to-engineer Sulfolobus islandicus rod-shaped virus 2 (Mayo-Muñoz et al., 2018). Only viral particles that...Editing of Sulfolobus islandicus Rod-Shaped Virus 2. Viruses 10:695 . https://doi.org/10.3390/v10120695...Bondy-Denomy et al., 2018). For example, AcrIIA4 mentioned above indicates that it’s the fourth Acr protein discovered...Cas9 Variant for Your Next Experiment Read more about Optogenetic Control of CRISPR   Resources on Addgene.org...

...Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system." Cell (2015). PubMed PMID: 26422227... Find plasmids from this publication at Addgene. 2. Kleinstiver, Benjamin P., et al. "Genome-wide specificities...Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell. 2015 Sep 23. pii: S0092-8674...a cell culture model. What we still don't know about Cpf1 In their original paper characterizing Cpf1...Addgene Blog Catch Up On Your Cpf1 Background Read About CRISPR Competitor NgAgo Learn How You Can Use piggyBac...

...groups, such as their vaccine production, SARS-CoV-2 research, or adenovirus packaging cell lines dataset...-Line Knowledge Resource. J Biomol Tech. 2018; 29(2): 25-38.... of Bioinformatics reached out to us to tell us about a resource called Cellosaurus, a growing cell line...all the information in the resource.  This is a labor-intensive project; scripts can be used in some cases...preferences to help you identify proximal lines you care about. If you have cell lines without a certified origin... Or finding errors in the available information about a cell line you’ve used? Submit your manuscript ...distributor. A success all around! If you’re excited about this resource and want to learn more, check out ...

...constructs for tagged and untagged expression of SARS-CoV-2 proteins in bacteria, yeast, and mammalian cells. ...approximately 3,000 constructs to support SARS-CoV-2 research.  Browse the entire Ginkgo Bioworks COVID...to COVID-19 Learn more about the Ginkgo COVID-19 plasmid collection Read about plasmids in the COVID-19... are showing an unprecedented amount of support about allowing use of plasmids for research by anyone....

...literature? Very often the data of interest are in 1 or 2 figures from one paper and then another figure in ...ReFigure by creating ReFigures and answering this 2 minute survey! You can also find ReFigure on Twitter...ReFigure Analyzing replications as in this ReFigure about whether Natronobacterium gregoryi Argonaute has ...functionality Collecting preclinical and clinical data about a disease or therapeutic Side-by-side comparison...share your science at We Share Science Learn more about Data Sharing Resources at Addgene.org Deposit ...

... is also gaining a lot of new traction lately (1, 2), depends on knowledge of the structure(s) a given...per plate) across 20 fresh selective plates. Step 2: Express random phage DNA fragments in E. coli Now...microbiology 23.4 (2015): 185-191. PubMed PMID: 25708933. 2. Cooper, Callum J., Mohammadali Khan Mirzaei, and ... biology 34 (2015): 69-77. PubMed PMID: 26275959. 2. Weigele, Peter, and Elisabeth A. Raleigh. "Biosynthesis...our planet, but we still know collectively little about how they infect and influence the evolution of their...

...initial work was with TALENs, in collaboration with the Porteus lab [2]. With the advent of CRISPR/Cas9...nucleases. Genome Res. 2010 Jan;20(1):81-89. PubMed. 2. Bauer DE, Kamran SC, Lessard S, Xu J, Fujiwara Y,...PhD student at Harvard Medical School in the laboratories of Stuart Orkin and Daniel Bauer. He received...

...) 38 kb Cells HEK-293T Sf9 Time to collection 2–4 days 5–10 days Purification Ultracentrifugation...Centrifugation Titration time ~1 day (AAV); ~1 week (lenti) 2–10 days Initial titers Higher Lower Resources... a factory to produce adeno-associated virus type 2 vectors. Human Gene Therapy, 13(16), 1935–43. https... widely available, so you’ll only have to worry about cloning your transfer plasmid. The transfer plasmids...typical insect cell preparations, which we talk about below. Processing The viral supernatant is typically...available, so the amount of cloning required is about the same between insect and mammalian systems. Baculovirus...dead cells and cellular debris. While adding time (about three to four days), this second round provides ...

...the result in Figure 1 could look like this: Figure 2: Expression of Plasmid B (as internal control) This...down expression of Gene X but, as shown in Figure 2, this was likely due to the original transfection ...PMID: 20074371. PubMed Central PMCID: PMC2817684. 2. Replicates and repeats—what is the difference and...optimization within your experiment. Read on to learn more about the various controls that can be used for plasmid-based...: Expression Level of Gene X The results shown above are from a single experiment wherein Plasmid A (...be compared. The Empty Vector Control (mentioned above) could also serve as an important negative control...experimental cells. For example, in the experiment above, Plasmid A contains an shRNA that targets human ...

... qPCR is that the results can vary by a factor of 2. This means that if you set up two identical assays...amplitude measured for each droplet in each well (Figure 2). A clean ddPCR should have a clear separation between...as low as 1C/s. Increasing the elongation time to 2 minutes and the denaturation time to 1 minute has ... droplet generator, you will have to be careful about transferring your droplets from the droplet generator...Additional resources on the Addgene blog Read more about polymerase chain reaction Browse our blog posts ...collection of viral vector related protocols Learn more about the Addgene viral service ...

... Plant Methods. BioMed Central, 2(1), p. 19. doi: 10.1186/1746-4811-2-19. PubMed PMID: 17087829. PubMed...PMID: 24032037. PubMed Central PMCID: PMC3759027. 2 Harrison, S. J. et al. (2006) ‘A rapid and robust ... J. (2002) Arabidopsis : a laboratory manual. Cold Spring Harbor Laboratory Press. Available at: https...https://books.google.at/books/about/Arabidopsis.html?id=IfZAMNPWVk4C&redir_esc=y (Accessed: 21 September ...

...experiment using smFPs and standard neuronal tracers. 2. Visualization of neuronal sub-cellular structures...similar proteins. For example, N-cadherin (cadherin-2) is a postsynaptic cell adhesion protein that plays...PMID: 25915120. PubMed Central PMCID: PMC4573404. 2. Shepherd, G.M. (2004). In "The synaptic organization...enable accurate labeling of neurons. As discussed above, weakly expressed proteins in particular are difficult...resembles images of the deity (see comparison image above). The church of the Flying Spaghetti Monster is ...tags. 4. High-resolutions microscopy As indicated above, smFPs can be used with conventional microscopes...

...synaptobrevin 2; At = ALFA-tag; BACE = β-secretase; AF = AMPA.FingR; FingR = fibronectin intrabody generated...RNA guide (known as an omega RNA, or ωRNA) (Figure 2), but TnpB is small enough to be carried by viral ...than a third of the size of Cas9). Figure 2: Comparison of TnpB (A) and Cas9 (B) genome editors...Check out our blog post on AAV serotypes for more about the tissue tropism of these and other AAVs. ...

...handled at BSL-1, while lentivirus requires a BSL-2 setup. This requires a less involved setup when working...infection rates in your experiment. Figure 2: Visual representation of common AAV serotypes used..., 10(1). https://doi.org/10.1038/s41467-019-12449-2 Pupo, A., Fernández, A., Low, S. H., François, A.,...Hanlon, K. S., Kleinstiver, B. P., Garcia, S. P., Zaborowski, M. P., Volak, A., Spirig, S. E., Muller, A.,...vivoscreening platform. bioRxiv (Cold Spring Harbor Laboratory). https://doi.org/10.1101/2023.06.13.544831 Santinha...

...properly exiting the ER and fluorescing (see Figure 2) [5].  When using FPs containing two cysteine residues...compartments including the secretory pathway (Figure 2), the inner membrane space of mitochondria and chloroplasts...PMID: 18228466. Pubmed Central PMCID: PMC2875081. 2. Snapp, Erik Lee. "Fluorescent proteins: a cell biologist's...moxFPs over standard FPs for the reasons listed above. In addition, because the moxFPs are highly monomeric...Medicine. Her doctoral research was conducted in the laboratory of Erik Lee Snapp and focused on optimizing fluorescent...

...upstream of an opsin gene (e.g., channelrhodopsin-2, ChR2) fused to a reporter gene (e.g., mCherry) to...the correct orientation to be transcribed (Figure 2) (Sharma and Zhu, 2014). How does FLEx enable strong... Abdallah, Khaled, et al. "Adeno-associated virus 2/9 delivery of Cre recombinase in mouse primary afferents...Deniz, et al. "A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range ...FLEx enters the scene. FLEx: A light when night is about you! The FLEx (for flip-excision) switch (aka DIO...PMC4166436. Resources on the Addgene blog Learn more about FLEx vectors Download our Plasmids 101 eBook Read...

...the reaction’s metabolites. One useful genetically encoded H2O2 probe is pQE-30-HyPer3 (2) which he will...Pharmacology 2017 Jun 15;134:1-17. PubMed PMID: 27984002. 2. Bilan, Dmitry S. et al. “HyPer-3: A Genetically Encoded...The Addgene-Seeding Labs plasmid grant is a collaborative endeavor providing plasmids to researchers in... of Zimbabwe, Zimbabwe Dr. Idah Sithole-Niang (above, left) is an Full Professor in Biochemistry with... CRISPR effect observed in bacteria. Learn more about the different CRISPR applications in plants.     ...Rodrigo Arturo Rivera Solís, Associate Professor (above, far right), thrives to make affordable biosensors... to the Addgene blog for updates and future collaborations. Many thanks to all of the awardees, all who...

... (1) the struggle between two opposing forces or (2) the result of competing desires or (3) the presence... an introduction, and a resolution.   Fig. 2: The structure of a story - not just for fiction! ... your science with others.    My favorite quote about science ever comes from an interview in which someone...storytelling business like to say, science is all about navigating conflict. In literary terms, conflict...find that once I sit down and talk to a scientist about their research, these conflicts come out quite naturally... a simple question, like “Have you ever thought about how X works?”, or even by introducing a person that...devices, we sometimes forget the most important thing about science: It’s cool. Like bowties! Using science ...

...the team (Fig. 2). These images are based on the concept of bacteriographs created about a decade ago. ...spatially precise the activation is.   Figure 2: A bacteriograph of Michelangelo's "The Creation of...Twitter, you might have seen the video of her singing about the paper recently published by her lab and Mustafa...professor at the University of Freiburg. This paper is about a new tool called BLADE (blue light-inducible AraC....   Inspired by @UriAlonWeizmann, here my song about our paper recently published in @nchembio Title:...Additional resources on the Addgene blog Learn more about inducible promoters Visit the Plasmids topic overview...

...resistance to the herbicide chlorsulfuron (Homo line 2) or wild type (WT), two weeks after herbicide was ... CLIPf tag and a C-terminal SNAPf tag; see Figure 2), the authors developed a cost-effective expression... when associated with microtubules. Figure 2: CLIP-SNAP fusion proteins for single-molecule FRET...synaptic events By Brian O’Neill The Lin Lab and collaborators recently created ASAP5, a faster and more responsive...

... flanking sequences to their input string (Figure 2). This string can then be used as input for the prediction...top 5) and test them in their labs. Figure 2: Guidelines to predict pegRNA efficiencies with pridict.it...Biotechnology. https://doi.org/10.1038/s41587-024-02268-2 Schep, R., Brinkman, E. K., Leemans, C., Vergara, ... al., 2023). We trained our models in close collaboration with the Krauthammer lab (University of Zurich...

...Epub 2016 May 2. PubMed PMID: 27136078. Find plasmids from this publication at Addgene. 2. Swarts, Daan...impaired NgAgo activity, with three mismatches abolishing activity. One-guide faithful NgAgo guide loading... how it’s working in the comments section! Read About NgAgo User's First Impressions Here References ... Additional Resources on the Addgene Blog Read About Cas9 Homolog Cpf1 Check Out High Fidelty Cas9 Mutants...

...indicating primer dimers. (c) A double peak indicating 2 amplicons. Image credits (respectively): Zuzanna K...details for reproducibility’s sake!   Fig. 2: Setting up triplicates in multiple configurations...used method is the Standard Curve method. See Table 2, (adapted from Wong, 2018) for a comparison of different...Statistically defined log-linear phase   Table 2: A comparison of different qPCR analysis methods. ... for RNA quantitation. Biotechniques. 2000 Aug;29(2):332-7. doi: 10.2144/00292rv02. PMID: 10948434.Taylor... should have a melting temperature 5-10 degrees above the primer and be at least 25 bp away from each ...

...identifying the resources needed to accomplish that goal. 2. Identify the tools in that timeline Avoid writing...based on experimental timelines and specific aims. 2. Then make decisions based on financial values. All...Would I be able to get access to these materials?” 2. Always negotiate for a win-win decision An institute...a successful negotiation and collaboration. We’d love to hear more about your experiences and opinions...can get away with using computer modeling and collaborations rather than using animals, then that’s a cost...associated with old equipment, like service repairs, labour costs, and loss of time, can exceed the cost of...Everything is negotiable. Try asking for discounts, collaborations, access to internal award applications, etc...

...into the several steps outlined below and in Figure 2. For a deeper dive into the steps and techniques, ...mounting sample on slides, and confocal   Figure 2: High-level overview of an indirect immunofluorescence...recognize the species of your primary antibody and 2) is conjugated to a fluorophore that suits your experiment... Learn about research applications of antibodies in Introduction to Antibodies Learn more about fluorescence...Stay tuned for a future blog post to learn more about these techniques!  Direct vs. indirect immunofluorescence...different approaches to be familiar with when talking about IF, direct and indirect. For direct IF, researchers...immunofluorescence procedure with key considerations about each step. Image created with BioRender.com   The first...

... Figure 2: Ironing boards. Good as a WFH desk, and as a video control center.   Cycle 2: Zooming ...ironing board, giving it the perfect height (Fig. 2). We recorded video using a simple, external video... Production Specialist, I’m constantly thinking about ways to improve our productions to deliver the best...four months after I started with Addgene, I was about to have a check-in with my boss Jennifer. I had ... would be a couple months. Jennifer and I spent about an hour that morning scrambling. Everything we had...someone to be comfortable on camera while thinking about things like lighting, camera angle, and shot composition... allowing people to sit without having to worry about getting in close to mic them up. Monitor: I then...