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...community submissions for Addgene and CLOVER AAV experiments - and we've updated our submission process ...say, should!) submit your data to the Data Hub! The process is easy with our new submission form - and no...step-by-step submission guide or email [email protected] to learn more. A great submission includes: ... cited by others?)  Ready to submit?  Fantastic! The next 25 submissions will receive a 20% off coupon...recording to see two reports presented by the submitting scientists, learn more about the AAV Data Hub...given by Connie Cepko and Ian Wickersham.     Submitting to the Data Hub If you've ever validated an Addgene...

...0.02–0.05 ml Skill required: beginner Subcutaneous Subcutaneous (SQ or SC) injections are given between...method, subcutaneous injections take some practice to get the coordination down. Subcutaneous injections...the mice, and restraint method. Many different substances can be injected, including cells, drugs, DNA,.... IV injections are used for a broad range of substances, as it circulate what is injected throughout ...technique. Intrathecal injections are used when the substance being administered must be delivered to the central...Intratracheal instillation is used to deliver substances directly to the lungs. It is an advanced technique...invasive, and only requires a pipette. Since the substances are administered through the nose, they also ...

...neuronal subtypes was not available until now. In order to target specific neuronal subtypes such as inhibitory...recently deposited 2 Bacillus subtilis libraries with Addgene – the B. subtilis Single Gene Deletion Library—Kanamycin...Every quarter we highlight a subset of the new plasmids in the repository through our hot plasmids articles...hot plasmids segment on the Addgene Podcast. Subscribe to the Addgene blog to get all the latest hot ...containing high fidelity Cas9 variant HF-Cas9. They subsequently tested the original BE3 and HF-BE3 using a gRNA...the podcast Segement on the pPSU Plasmids B. subtilis knockout and knockdown libraries The Gross Lab... Essential gene Knockdown library. Although B. subtilis has a fully sequenced genome, many applications...

...are not subject to photobleaching/phototoxicity, they successfully imaged multiple subcellular compartments...luminescent substrate is required for imaging, and it is important to verify that this substrate does not...fused to an FP; when supplied with a luciferase substrate, the luciferase transfers energy to the FP, resulting...excitation light source. Instead, a luciferase substrate like coelenterazine is supplied in the culture... which is limited by depletion of luminescent substrate, they synthesized diacetyl coelenterazine-h. Unlike...coelenterazine-h, producing a constant supply of substrate. Using this compound to excite the Nano-lanterns...brightest characterized luciferase, NanoLuc, and its substrate furimazine. In addition to the previously described...

...ligate many undesirable structures, including substrates containing gaps or DNA base pair mismatches (...such as Taq DNA Ligase, which only reacts with substrates containing no gaps, and will not join any fragments... sequence is present, the probes will ligate. Subsequent cycling through rounds of melting and annealing...generally prefer fully Watson-Crick base-paired dsDNA substrates to those containing one or more mismatches. However...pyrimidine mismatches are typically worse ligation substrates than pyrimidine:purine mismatches. Helix stability...crystal structures of several DNA ligases bound to substrate (23, 24). Thermostable DNA ligases, including ...naturally able to discriminate against ligating substrates containing base pair mismatches (i.e., are "higher...

... have subscriptions to the specialized journals in your field? Post a preprint before you submit to a ...article, presenting a conference poster or talk, or submitting patent or grant applications; to informal settings...especially the increasingly growing time from submission to publication. It takes many months, often more...the possibly repeated rounds of rejection and resubmission to other journals. Preprints allow authors to...admittedly, somewhat meaningless) practice of listing “Submitted” articles in their CV or grant applications. A...post a preprint at the same time as, or before, submitting the manuscript to a journal for formal peer review...opportunity to strengthen the manuscript prior to submission. Preprints at Addgene Preprints are all about...

...cytosine-to-thymine substitutions in target sequences (Banno et al., 2018). These substitutions were limited ...enable C-to-T and A-to-G base substitutions respectively. Substitutions seem most efficient when using...time needed for a single round of editing, and a subsequent curing protocol that requires only 2-3 hours ...days, and the pTargetF and pCas plasmids can be subsequently cured from the bacteria. Although developed ... the transcript. Different Cas orthologues and subtypes function somewhat differently and can be more ...Cas9 (dCas9) alone or a dCas9 fused to the omega subunit of RNA polymerase, the lab   successfully repressed...such a purpose (Selle et al., 2020).  Targeting a subset of bacteria in a population Work from the Lu lab...

..., it cannot be used for subcellular analysis such as morphology or subcellular localization studies. For...cell? Or perhaps, you need to analyze a specific subset of cells in a complex population. Have you spent...this technology can be used to isolate a specific subset of cells in a population in a procedure termed ...not, however, permeabilize cells and requires subsequent permeabilization with detergents such as triton-x...instruments have specialized components that can divert a subset of cells. During FACS, the sample stream oscillates...each well allowing users to easily isolate and subsequently expand monoclonal cultures.  Interpreting flow...they are interested in and further analyze that subset of cells. An immunologist, for example, might be...

...follows: Cap Subject Industry H1B Academic H1B National Interest Waiver EB-1A EB-1B Cap subject industry...companies to submit an application on their behalf.  If an individual obtains a cap subject H-1B (last ...numerically limited visas are referred to as “cap” subject H-1Bs. The H-1B allows U.S. employers to employ...numerical quota. Four types of employers are not subject to the H-1B cap: institutions of higher education...national’s proposed work endeavor must have both substantial merit and national importance; (2) The foreign...show that you serve the national interest to a substantially greater degree than the majority of your peers...Addgene Blog Transferable Skills Guide 6 Steps to Submitting a Resume That Gets Seen Tips for Writing a Good...

...combinations of different Gα, Gβ, and Gγ subunits. When inactive, these subunits are found in a trimeric complex...transducer complex consists of a trimer of three subunits: alpha, beta, and gamma (Gα, Gβ, and Gγ), collectively... the GPCR, leading to the activation of the Gα subunit. The activated Gα exchanges bound GDP with GTP,...GTP, resulting in the disassociation of the Gα subunit from the Gβγ dimer, activating downstream signaling.... With binding of the drug, the alpha and beta subunits dissociate resulting in decreased BRET2 signal... are then treated with the luminescent enzyme substrate coelenterazine, followed by the addition of the...

...and α. Some isotypes have different versions or subclasses that have minor differences in their heavy chain...the hinge region in IgM antibodies. IgM has no subclasses and is found mostly in blood. IgD While IgD is...monomer structure with a valency of two and has no subclasses. IgG IgG is the most abundant isotype in blood...system encounters  an antigen for a second or subsequent time. It is expressed as a monomer with a valency...valency of two. There are four IgG subclasses, numbered based on their abundance in blood: IgG1, IgG2, ...involved in allergic reactions. There are no IgE subclasses and they are expressed as monomers with a valency...dimer with a valency of four. There are two IgA subclasses: IgA1 and IgA2.  You can learn more about isotype...

.... Application submission  Set a calendar reminder for every deadline and shoot to submit your application...deadline. The day of submission, review your application with a fine comb and hit submit. Check your email...find internships and scholarships in different subject areas!    2. Resume and cover letter know-how...your network to polish the final draft prior to submission. When in doubt, consult the web to find the tools...email for confirmation of submission. If you do not receive such a notice within 24 hrs, send a note to the...

...antibodies contain a mix of different isotypes and subclasses. They each have their own advantages. Since monoclonal...species, like an anti-mouse IgG antibody. Some are subclass specific, like an anti-mouse IgG2a antibody. When...    Figure 1: Broadly reactive versus subclass-specific antibodies. Only a fraction of the antibody... antibody (A). To boost signal, consider using subclass specific secondary antibodies instead (B). ...antibody contains a mix of different isotypes and subclasses, use a broadly reactive secondary antibody to...antibody you may wonder, “Will the isotype, and subclass affect my experiment?” Well, it depends on the... to reduce off-target signals. Use isotype- or subtype-specific secondary antibodies when staining multiple...

...different species, or at least from different IgG subclasses, so that secondary antibodies exclusively recognize...Must use different species and/or different IgG subclasses of 1° for multi-color staining Can be expensive...limited cross-reactivity between different IgG subclasses and different species, nanobodies are useful ...require using primary antibodies from different IgG subclasses. Instead, you can pre-incubate individual primary...antibody: species it was raised in and its IgG subclass. Once you know the answer to these two questions...right nanobody for your work. Species Antibody Subclass Nanobody Addgene Plasmid Rabbit N/A* TP897... (3x cysteines) *Rabbits have only one IgG subclass.   Express the Nanobody In Bacteria: Table 2 also...

...incremental, ordered cell lysis. E. coli cells are subject to lysis by freeze-thaw if not suspended in cryoprotectant...

...against the cellular ribonuclease P/MRP 30 kDa subunit (RPP30) are used as a control for normalization...

...encourage you to submit data showing how the antibody worked in your application. The submission process is ...Antibody Data Hub. Data reports  Each antibody submission is used to create a data report, which contains...data showing how the antibody performed in the submitted application, along with a brief description of...reports Just like our AAV Data Hub, each data submission we receive is checked by Addgene's curation team...so you can work through it at your own pace. To submit, simply click "Contribute Data."    Figure 6: ...

...s do it!'" Working as a subject matter expert Caitlin started out as a subject matter expert (SME) at ...more about Caitlin Runne-Janczy and her job as a Subject Matter Expert/ Product Development Manager at eScience...is often like being a jack-of-all-trades. “As a subject matter expert in the educational sphere, it can...it is researching the different aspects of the subject you're writing and then condensing that and making...Hutchinson Cancer Research Center. And make sure to subscribe to the Career Channel of the blog so you don’t...

...serotype did you use and what animal subject did you use? For animal subjects it is helpful to report the genetic...location, titer dose (Example: 1E13 GC/mL) and subject age at time of injection. How long post-injection.... Image: The final and required step requires submission of an experimental image result that you generated...this provides a clear overview of what you need to submit data. Please note that the data hub is currently...viewing or data deposition experience. If after submitting or viewing the data you have any feedback at ...

...if not for this specific submission, then for the submission to the subsequent journal.  As a postdoctoral...Saudi Arabia. From the perspective of an author submitting a paper, the peer-review seems like another dragon...tolerance, I have been peer-reviewing journal submissions since 2015 for a wide variety of journals. Since...routine that works well for me to evaluate the submitted paper in addition to the journal-specific guidelines...