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...that screen with a system you are really confident about working for most genes and individual targeting ... system, Konermann explained, was new knowledge about Cas9’s crystal structure. “I thought maybe we could...RNA we use. What we have to do now is learn more about the rules.” He said that there is no theoretical...References & Further Reading: For more information about SAM, including a tool for getting optimized guide...

...transitioned away from the bench and became all about finding the best product-market fit for this set...Notably, the team must have an upfront conversation about its goals and vision for the company; this will ...section! And read on for more great information about starting a biotech company. Suggested reading The...of Sample6) for taking the time to speak with us about the biotech startup process and work environment...

...cloning before you can start your experiment? What about all the reagents you need to order? Sometimes, it...dive deep into generating the data you’re excited about. When we were closing in on the one million plasmids...million plasmids shared than not!), we think it’s about time we updated this post. How much time does ...choose the right cloning method Find blog posts about scientific sharing here Download the Plasmids 101...

...fragment are we talking about here? Many of the Fab fragments described above can be used for similar ...fragments are produced by proteolytic digestion above the hinge-region, which results in two independent...you consider which tools to use, be sure to think about what pieces of the antibody you do and don’t need...

...August 2nd in Nature gives us even more to talk about, as it represents another step towards CRISPR germline...impossible with current tools, thus quieting some fears about the creation of “designer babies.” The differences...of individuals with varied backgrounds. Talking about this study with non-biologist friends Like me, you... Blog Check out Our New CRISPR Topic Page Read about High Fidelity CRISPR Variants Target Multiple Genes...

...source of monoclonal antibodies, but they are laborious to generate and costly to share. What is a RANbody... particular applications. Read on to learn more about each reporter or check out table 1 for a summary...Additional Resources on the Addgene Blog Learn more about using the secondary nanobody toolbox for immunodetection...can be found here Watch this video to learn more about nanobodies Additional Resources Looking for primary...

..., social media, coffee chats with potential collaborators, blogging and more. These each have different...key routes for researchers to share information about the emerging COVID-19 pandemic quickly and openly...awareness of the benefits of preprints. Some worry about faulty work being posted as preprints, assuming ...submission. Preprints at Addgene Preprints are all about sharing scientific knowledge faster and more openly...

... blog Read more about the PHP.eB AAV capsid Read more about optogenetics Learn about the differences ...library were designed using structural information about parental channelrhodopsins to create recombinant...

.... Gendered Innovations is an international, collaborative project—funded by the European Commission, the...research.  In addition to the studies referenced above, another excellent resource for research design ... whole human population. To learn a little more about where differences in effects on the sexes come from...cost. Especially in animal research, they worry about needing more study subjects and control groups. ...

...One of the best things about sharing plasmids through Addgene is that we provide an added level of confidence...bp of clean sequence from a typical reaction, or about 250 amino acids worth, so make sure the feature ...96-well microplates (identical copies, see image above). We then select forward and reverse primers for...specifically to target this region, as outlined above. Avoid primers that bind within or must read through...

...Nielson and Voigt (2014).   We talked with Voigt about this collision that’s taking place between CRISPR...straightforward surprisingly. So I think a nice thing about CRISPR is how much easier it is to work with than...can slow their growth. Something has to be done about that.  The second problem is fundamental. We built...worked in each case. Sometimes in CRISPR people talk about off-target interactions. That’s relevant if you ...

...a broad range of reproductive, developmental, metabolic, and immune response programs. NRs also encompass...gallstone disease, senescent illnesses, and the metabolic syndrome. Coregulators Coregulators constitute... regulatory regime (such as genes involved in metabolism, growth or cytokine action). Primary or core ...and the free exchange of these vectors between laboratories. The development of the transient transfection...

...CRISPR Featured Topic Page Learn more about Cpf1 Find out about CRISPR Multiplexing Resources on Addgene.org...coli and other bacterial species. Read on to learn about the tools available for bacteria and some of the...that can be used in this way. As described in the above CRISPRi section, the Marraffini Lab published a ...as a collection of organisms, their genes, and metabolic processes, occupying the gut. Many human diseases...

...can be done using FRET! Read on to find out more about this amazing imaging technique and find further ...our curated list of biosensors too, as another laboratory may have already created the sensor that you ...ability of labs to answer sophisticated questions about cellular processes. Currently, FRET experiments ... Tips for using FRET in Your Experiments Learn about Fluorophore Targeting for FRET Studies FRET Based...

...blog Learn about the advantages and disadvantages of using monoclonal antibodies. Read about other techniques... require two different antibodies. As mentioned above, it can also be used to characterize antibodies ...concentration of proteins in an ELISA well As mentioned above, the measurement of the concentration of proteins...

...you do to get ready for this interview? Maybe the above scenario sounds familiar to you. It was my experience...positions often required a one-hour presentation about a prior research project. In total, the interviews... find lists that have lots of general questions about job responsibilities, performance evaluations, professional...require. Frequently they will ask you to present about your prior research. This is a chance to highlight...

...general consideration for behavioral testing in a laboratory, a high level of experimental control is required... after stereotaxic injection. Find more details about choosing AAV or lentiviral vectors in our previous...it’s crucial to appropriately time the injection about two weeks before the behavior test. This time window... Viral Vectors Resources on Addgene.org Learn about Addgene's viral service Find viral protocols and...

...purposes of labor laws, rather than as trainees in a special category, as has been the ‘labor-of-love’ philosophy...passing and upcoming implementation of the Federal Labor Standards Act (FLSA) which mandates that all workers...event of note is a ruling in August by the National Labor Relations Board to allow graduate students who work...

... are nice if you can afford them). I am talking about making sure the audience is interested, educated...of the stories or topics you know they can speak about. There should be some spontaneity, but having a ...location and parking information and remind them about the event as it approaches. Room set-up and logistics... make or break the discussion section. And what about those audience members that want to take over? It...

...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...with the firmer 2% agarose gel solution discussed above for picking. Many thanks to our guest blogger Iris...

...the context of the cell. However, much knowledge about functionality can be gained by understanding the...subcellular marker proteins, it’s possible to get an idea about the subcellular structures your protein resides ...with. These “colocalization” studies give insights about the proximity of two proteins within the same subcellular...Additional Resources on the Addgene Blog Learn More about Plasmids for Endogenous Gene Tagging in Human Cells...

...of the hood while work is in progress.Learn more about cell culture contamination in this post from the... bath constantly at 37°C for this purpose.Learn about the effects of CO2 on sample pH Cell morphology:... Set up a routine to split cells Don’t be lazy about splitting cells, instead try to form a routine. ...secretion of lactic acid (a by-product of cellular metabolism that is toxic to the cells), and need to be split...

...If you have ever worked in a molecular biology laboratory you have likely done a polymerase chain reaction...uses ddPCR for AAV titrating. You can learn all about ddPCR for AAV titration and find helpful tips and...method that can help save time and effort in the laboratory.  There are two main categories of multiplex ...use a single pair of primers, like those stated above. Multiplex PCR is also used in high throughput SNP...

...technology. To dig more into what researchers felt about NgAgo, I conducted a survey and asked researchers...successful, and the second section asked basic questions about the NgAgo protocols used by respondents (transfection...27136078. 2. Burgess, Shawn., et al. "Questions about NgAgo." Proteins & Cell (2016). 3. "NgAgo: Genome-editing...PMC4343198. Resources on the Addgene Blog Learn More about NgAgo Enhance CRISPR Targeting Specificity with ...

...directly back to your specific goal. When thinking about step 3, however, you may be surprised at the number...Delivery methods for mammalian cell lines As mentioned above, the expression system you choose in many ways dictates.... The user should review the current literature about their preferred model.  If your expression system... Check out our CRISPR Featured Topic Page Learn about CRISPR Delivery Using Ribonucleoproteins (RNPs) ...

...genes above with your gene or genes of interest. Some of the plasmids listed in the table above (and their... an internal region of the mRNA. In the figure above, you can see that the IRES element acts as another... first discovered in picornaviruses, are short (about 20 amino acids) and produce equimolar levels of ...

...CRISPR Featured Topic Page Learn about Non-Homologous End Joining Learn about Base Editing Read 3 Tips to ... post on increasing HDR efficiency to read more about optimizing this distance. Homology arms and types...screening strategies for your experiment. Also learn about Addgene’s knock-in plasmids for reagents to help...

...Read this blog post about CRISPR Kinome Libraries available at Addgene Learn about the advantages of pooled...large-scale functional screens Read this blog post about genome-wide screening using CRISPR Resources on...libraries for the mouse genome! Other Resources Read about CRISPRi and CRISPRa from the NIH E-newsletter ...

...gene therapy applications. Read on to learn more about the current (and future) applications of lentiviral...do both, Addgene depositor Didier Trono and collaborators turned to the lentivirus HIV-1, well-known to...Libraries Enable Genome-Scale, Knockout Screening Read about Genome-Wide Screening Using CRISPR/Cas9 Additional...Addgene.org Check out Our Lentivirus Guide Learn about Virus Biosafety Find Lentiviral Vectors for Your...

...genome editing than Cas9 - read on to learn more about Cpf1 and check out our CRISPR guide for a refresher...mediated cleavage are 42-44 nucleotides in length, about the same size as Cas9’s, but with the direct repeat... Cpf1 was first identified, we didn't know much about its on-target and off-target editing efficiency....CRISPR system proved we had a lot more to learn about CRISPR biology. Later work, like the adaptation ...