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...training progresses.  In From Academic Solos to Industrial Symphonies, an article well worth reading on ...can directly interfere with a seamless jump into industry.” In his article On Leaving Academe, Terran Lane...excellent—some say “cushy”—tenured faculty position for the grind of corporate life”.   In fact, there are a lot of...budget constraints Academic science is much like industry but the “product” is publications—the pressure...—science can be surprisingly more rigorous in industrial settings Tight (sometimes impossible) timelines—others...training and resources within the company Must find own sources of knowledge often externally A bit...

...not overgrow the plasmid at 37°C. Some researchers find that using a 30°C incubation temperature to grow...vector backbone vs. the full length plasmid. If you find that your bacterial cultures contain a mix of full... bands: nicked, linear, and supercoiled. Here we find a smaller, recombined plasmid around 1.5 kb. When...the recombined plasmid is still visible. This indicates that the DNA prep contains a mixture of the full....069286.118 Additional resources on the Addgene blog Find more molecular biology and viral vector protocols...protocols Learn about the common lab E. coli strains Find tips for viral transduction  ...

...Allele Biotechnology, Florida State University, Indiana University, and the Karolinska Institute developed...immunoprecipitation experiments with other fluorescent proteins. Find mNeonGreen plasmids here! mTFP1: a monomeric teal...insensitive to physiologically relevant changes in pH. Find mTFP1 plasmids here! mWasabi: a bright green fluorescent... significantly excite at 400 nm while EGFP does. Find mWasabi plasmids here! One of the most notable things...Addgene blog Read our fluorescent protein blog posts Find blog posts about choosing fluorescent proteins for...Addgene’s fluorescent protein plasmids and resources Find fluorescent protein kits ...

...Resources on Addgene.org Find molecular biology educational resources Find protocol videos to teach basic... and understand how therapeutic strategies work. Find Addgene Blog posts about other educational tools...Accessing and modifying BUILD YOUR PLASMID You can find rules, suggestions, and more details on how to play...FRE_SCI.   Additional Resources on the Addgene Blog Find planarian tools for teaching developmental biology... can help teach students about Synthetic Biology Find Arabidopsis educational resources at the Arabidopsis...

...Additional Resources on Addgene.org Find Ready-to-Use AAV Preps Find Lab Protocols for Work with Virus ...titers Making contact with a cell Although an individual AAV particle’s ability to infect a cell is dependent...concentration. In this flowchart, you can see the dwindling proportion of AAV particles that make it through...high titers, as contact between the cell and any individual AAV particle is rare. But simply getting into...higher titer you will have. When possible, use low-bind or siliconized tubes to store samples as it is more...

... at 230 nm. The A260/A280 ratio is used as an indicator of DNA purity. Ideally, this number should be ..., you can use the agarose gel method to not only find out how much DNA you have, but also to see whether...like a PCR product). This method also gives an indicator of DNA or RNA contamination based on the presence...no other instruments are available. Some last reminders about quantifying DNA In choosing your DNA quantification... more about molecular biology protocols and tips Find the best way to elute and store your plasmid DNA...Addgene.org Visit the Molecular Biology Reference Find plasmids for your research Browse protocols for ...

... previously. While this claim has yet to be independently verified, the watching world erupted in controversy...private funder, government regulators, and an independent ethics council – all worked in tandem, within...experiment on humans, and they are not remembered kindly. However, despite many headlines questioning if...utilizes her firsthand experience in the software industry to inform her work on emerging issues in novel...theirpenmanship. Additional resources on the Addgene blog Find all of our CRISPR blog posts Download the CRISPR... Resources on Addgene.org Read our CRISPR guide Find CRISPR plasmids by function Check out our genome...

... phosphorylates a response regulator (RR). TCS-induced responses are varied; one outcome is directed transcription...promoter strength of various components, removing inducible promoters that could crosstalk with other pathways...doi: 10.1021/sb500273n. Epub 2014 Oct 11. PubMed. Find the plasmids from this publication at Addgene. ...CA. Nature. 2005 Nov 24;438(7067):441-2. PubMed. Find the plasmids from this publication at Addgene. ...1272-81. doi: 10.1016/j.cell.2009.04.048. PubMed. Find the plasmids from this publication at Addgene. ...10.1016/j.jmb.2010.10.038. Epub 2010 Oct 28. PubMed. Find the plasmids from this publication at Addgene. ...

...synthetic beads that can bind to a conjugated antibody and beads that are unable to bind to them: like cells...real results from background noise or nonspecific binding. Flow specific controls The three types of flow...they are non-specific. The compensation beads can bind to any primary antibody you add. Single color controls...allow you to determine what level of non-specific binding you have in your sample. The isotype antibody will... This enables you to identify any non-specific binding to know that your positive cells are truly positive...

... and Immunity and learn more about “... recent findings in basic immunology, cancer-immune cell interactions...discussed the effects of starvation on cancer therapy. Findings in previous animal studies suggested that fasting...in trying to resolve the inflammatory state by inducing immunosuppressive measures, for example the upregulation...impressive videos of these interactions (Ref 3). You can find useful tips on in vivo imaging in our Fluorescent...scientific community, contact [email protected] to find out more about our free deposit and distribution...Browse Cancer Plasmids Browse Immunology Plasmids Find Kits for Cancer Research  ...

... an individual to another individual perspective? A: CAP-B10 and CAP-B22 were tested individually in 4...question that didn’t get answered live - but he kindly took some time to address all your unanswered questions... 4 marmosets each. Brain potency for each individual is reported in Figure 4B (see left for image and ...a first time AAV user Resources on Addgene.org Find Adenovirus Plasmids Adenovirus Guide Page Subscribe...

...able to find strategies and systems that set you up for success in the detail-driven daily grind of science...tedious, but it also works, so do it anyway.  You may find that after running a protocol a few times, you no... blot protocol, from a full page to a few key reminders on a sticky note   Visual Planning  Plate ... use cold block systems or buckets of ice. Try finding ways to intentionally set up your bench to help...forth, continuing all day or even all week. By finding ways to intentionally slow down, you’re giving ...

... passed down through cell division. You can find all kinds of different lentiviral and AAV plasmids as...protocols for plasmid cloning Find our molecular biology reference here Find molecular biology protocols... of a fine needle to deliver nucleic acids to individual cells. Electroporation on the other hand uses...edition (2000). Meibom, Karin L., et al. "Chitin induces natural competence in Vibrio cholerae." Science...

...of interest, as it is prone to generating indel errors. Indel errors generated in the course of repair ...where mutations are introduced. Repair of Cas9-induced breaks by NHEJ As illustrated below, NHEJ-mediated...microhomologies”). NHEJ doesn’t obligatorily introduce indels. Given the end structure of the Cas9 DSB (blunt... of accurate repair are easily re-cleaved while indel products aren’t, so repeated cycles will favor accumulation...constitutively expressing a targeted Cas9 should possess indels in the majority of their chromosomes within a day...

... at multiple levels to generate anything from individual transcription units (TUs) used to express your...metabolic pathways. For many MoClo systems, the individual parts are reusable, saving time and money on ...some common MoClo applications and kits. Keep in mind that this technology is useful for a variety of ...purification tags, connectors, cDNAs, and more to find the perfect combination. The EcoFlex Kit has a wide...array of DNA parts that can be assembled into individual TUs or very large (up to 20!) multi-TU constructs...et al., 2016). The variety of constitutive and inducible promoters, linkers, and tags makes EcoFlex a great...kits are designed with specific model organisms in mind, others are designed for flexibility. The Fragmid...

...established, it’s also a process that’s difficult for individual labs to complete. The nanobody based RANbody ...purification. Antibodies are difficult for an individual lab to produce - Hybridomas offer an alternative...RFP,  the histone protein H2A2B, and the actin-binding protein gelsolin, but any nanobody sequence could...serves as a backup means of detection that’s independent of the reporter. The only caveat is that HA staining.... Staining with multiple HRP-RANbodies is indistinguishable since they produce the same signal, so only...plasmids from Yamagata et al can be found here. Can’t find a RANbody against your favorite protein? No worries...

...system, antibodies are produced by B cells. Each individual B cell produces antibodies that all recognize...purification is one common method. These proteins bind the IgG heavy chain. Running the serum over a column...column of resin crosslinked to protein A /G will bind the IgG antibodies present in the serum, whether... are isolated from the serum by their ability to bind the target antigen. In this case, the serum is run...antibody; the ability of polyclonal antibodies to bind many epitopes of their target protein thereby greatly...polyclonal isotype control, to account for nonspecific binding. Now that you understand the pros and cons of choosing...

...2013). We found that the fluorescent proteins can, indeed, interfere with the correct function of the protein... species (bottom panel in Fig. 1). Our results indicate that the tag protein may have an important influence...experiment. Since the fused protein could represent a hindrance to the successful expression of the protein of...opsin-expressing cells). For other things to keep in mind when making fluorescent protein fusions, check out... posts Read about chemogenetics vs. optogenetics Find more about the many types of protein tags References...Addgene.org Check out Addgene's Chemogenetics Guide Find fluorescent proteins at Addgene ...

...qualify what kind of “plate” you mean. Without context, it’s difficult to know what kind of plate they...figure out the sources of miscommunication and b) find practical solutions.  My PhD prepared me remarkably...document, I update the training frequently. In hindsight, getting a list of frequently used words like ...Addgene blog Read about other transferable skills Find all of our science career related blog posts Read...communication blog posts Resources on Addgene.org Find careers at Addgene ...

...next week!”, and “This is his job!” filled my naive mind. Moving forward from that first experience, I learned...process, she has faith in you! Additionally, keep in mind that universities require application submission...the mission of the agency and benefits the field reminds the reviewer why your project is the project to...travel, etc. As a first time budget writer, you may find this section challenging because it’s hard to determine...students and postdocs will be hired, direct and indirect costs, etc. It may be helpful to ask your advisor...application process? What types of writing feedback do you find most helpful? Have a great grant writing tip? Share...