...more CRISPR tools are being published each month, making CRISPR a great choice for your next experiment!...control gene expression. Base editors can help you make precise point mutations near the gRNA target site...single-stranded DNA oligonucleotide, so you will need to make sure your expression system and delivery methods...Envelope plasmids provide the necessary components to make lentiviral particles. Stable, tunable expression...

...applications, and the quality of a standard curve can make or break an experiment. Here, we will provide an...the concentration of the unknown sample.  How do I make the standards for a standard curve? You create a...with BioRender.com.   When choosing a standard, make sure that the sample is pure and free of any contaminants...start with a higher concentration of standard or make smaller dilutions.        As mentioned above, it...

...Science: What the Future Holds and the Scientists Making It Happen, is an exercise in restraint. The very...topics without some editorializing, and he wisely makes no such attempt. He is transparent about his own...be shared by fellow scientists--but that doesn’t make his introduction to these topics any less effective... revelations. The issues he discusses have been making the rounds in journal editorials, science social...

...messy, so you want to make sure you have sufficient volume to dip all your plants. Make sure your Agrobacterium...agrobacterium: Getting a transformant takes a long time. Make sure the agrobacterial culture you are growing definitely...increase background growth of non-transformants and make finding true transformants more difficult. Don’...

...chemists and biochemists have developed techniques to make labeling chemistry easier and more functional in...and continual improvement of fluorescent proteins makes them the go-to choice for many (if not most) fluorescence...labeling strategies and improved fluorophores are making chemical dyes increasingly attractive and accessible...Medical Institute. He would rather be in the lab making new fluorophores for advanced imaging modalities...

...lab-use tablet? The software also makes it easy to annotate protocols, make them public for others to use,...of useful molecular cloning protocols, including making LB agar plates, restriction digest of plasmid DNA...bench, keep your inventory at your finger tips, and make shopping lists as you finish the last of a reagent...

...grumpiest of researchers can enjoy it at times! What makes it rewarding is when you can tell a trainee is understanding...to show up as your most prepared, can-do self and make the most of it! So, what can you do to build a ...reviewed above. Offer to help clean up and/or make necessary reagents for the experiments. Unlike in...need help with…now”. Ask for feedback so you can make adjustments. This could look like “What are some...

...pre-made viral preps. If you can’t (or don’t want to) make viral preps in your lab, there are custom viral ...verified!–your pooled library DNA, the next step is to make your virus. If the library happens to have a specific...vary with titer, cell type, and other factors, so make sure to put that aliquot to full use! Common conditions...chances that a cell doesn’t get infected at all. To make sure that every plasmid’s payload is adequately ...

...advent of CRISPR/Cas9 has made it easier than ever to make precise, targeted genome modifications. Cas9 has...many available through Addgene’s repository) and make it easy to find your perfect match. Choosing a ...xCas9 has a PAM requirement of NG, GAA, or GAT, making it a more flexible cutter with the added bonus ... You can achieve your precise edit without even making a break.  Base editors are dead Cas9 (dCas9) or...

...their initial assumption. Eugene Koonin and Kira Makarova, longtime collaborators of Omar and Jonathan and.... Using computational observations, Koonin and Makarova found that Cas7-11 appeared to have derived from...generation came Prime editing — the ability to now make any small changes, including base edits, insertions...Özcan A, Krajeski R, Ioannidi E, Lee B, Gardner A, Makarova KS, Koonin EV, Abudayyeh OO, Gootenberg JS. Programmable...

...nowadays, we cannot make it a full month without a new, drug-resistant ‘superbug’ making headlines in one... develop survival mechanisms. Some microbes even make their own antibiotic compounds. In such a case, ...resistant bacteria to over-take the population and making them more difficult to treat in the future. Patients...

... for talk is over and instead we must make real changes to make the system sustainable. For years, there...Department of Labor issued a Notice of Proposed Rulemaking, soliciting feedback by Sept 4th.  They proposed...opportunity to accept these recommendations, and make gradual adjustments towards this new level. Indeed...

...larger target, such as a protein. Such flexibility makes aptamers ideal candidates for biosensors. Aptamer... of how they actually “report” as biosensors. Making aptamers glow The flexibility of aptamers means...rather than separate them (Fig. 1b). One can also make “signal-off” biosensors in which the aptamers fluoresce...decrease background in vivo. The nature of aptamers makes them excellent candidates for developing assays ...

...aggregation and cross-react with anti-GFP antibodies, making them difficult to use in a multicolor/multichannel...fluorophore-labeled antibodies with very specific reactivities making them more useful for multicolor imaging experiments...Sample preparation for these techniques sometimes makes it more difficult to detect protein tags using antibodies...Blog Rabies and neuronal tracing Retrograde AAV: Making the journey from axon to nucleus AAVs CREATEd for...

...preference, some people like to make stricter gates while some like to make looser ones (Figure 3). A more...free to use SSC for a single cell gate. However, make sure to compare H and A (or W and A).   Live... type of stain whether a tighter or looser gate makes sense. Here, luckily, the vast majority of my cells...

...used for delivering your light stimulus. This can make or break your experiments!  First, closely read ....2021.778900 Mattis, J., Tye, K. M., Ferenczi, E. A., Ramakrishnan, C., O’Shea, D. J., Prakash, R., Gunaydin, L... B., Kadmon, J., Raja, C., Chibukhchyan, A., Ramakrishnan, C., Inoue, M., Shane, J. C., McKnight, D. J...science.aaw5202 Mattis, J., Tye, K. M., Ferenczi, E. A., Ramakrishnan, C., O’Shea, D. J., Prakash, R., Gunaydin, L...

...yielding 1-10% knock-in. Read on to learn how you can make CRISPR mouse model generation easi-er! Generating...months. But certain models remain more difficult to make. For example, large insertions, like a fluorescent... site. If you are using the ivTRT method, try to make the first two bases of the left homology arm guanine...activity. To avoid re-cutting by Cas9 after editing, make sure that the repair template does not contain the...

... of DNA as inert. It generally stays put, which makes it easy to locate in a genome. But there is a type...genomes of many other prokaryotes and eukaryotes, and make up ~45% of the human genome (Munoz-Lopez et al.,...considered replicative since every time they jump they make a copy of themselves. This increases the number ... by their nature, are mutagenic elements, which makes them a great tool for mutagenesis screens that detect...

...non-dividing cells, including stem cells. These properties make lentiviral vectors fantastic options for delivering...such as macrophages and microglia. This property makes the lentivirus an attractive choice for a gene transfer...the accessory factors in HIV positive cells could make the virus replication competent. The 3rd generation...transgene resulting in low inducibility and potentially making it difficult to interpret your results. This can...

...you get the idea. What if there were a way to just make one fusion protein that switches to any color or...experiment (Figure 3). And using self-labeling tags makes updating your labels easy! Instead of taking the...ligands is more expensive than letting the cells make a fluorescent protein for you. And some ligands ...Selecting Fluorescent Dyes and Ligands GCE4All: Making Genetic Code Expansion Accessible Addgene's Genetic...

...differentially expressed genes can be the next stage in making sense of such data. These targeted screens can ...you have your list of genes, Green Listed can help make the gRNA design process quick and easy. Green Listed...information that you have input in the web interface, making it convenient for you to later identify how you...letting us use their reference libraries thereby making it possible to put together this software. To me...

...you go!  Tips to Make the Most of a Scientific Conference Poster Session TipsHow to Make Friends and Meet...Specifically, Jo, Brook, and Angela are co-leading "Making a Greater Impact Using Clear and Accessible Scientific...

...target bases without breaking the DNA backbone, making them efficient and precise tools for altering DNA...the core ring structure intact, allowing them to make 4 out of the possible 12 base-to-base conversions...were engineering an ABE to reduce its tendency to make off-target edits to RNA. Like other ABEs, the editor...mammalian cells Researchers in the Xueli Zhang lab were making similar observations around the same time. (The...

...CVRHutchinson #SciArt #STEAM #WinterWonderlab @Addgene Make your own here!: https://t.co/AnltWfKuQg pic.twitter.com... wrapped gifts using life science catalog pages. Make your WFH space feel a little more festive this holiday... entering in (if you don’t include this, we will make our best guess). Follow @Addgene, use the hashtag...

...experiments. Best practices!). Making the find plasmids badge appear So how do you make this magical Addgene badge...favorite topic. If you have any other cool tricks to make PubMed browsing even easier and more useful, let...

...recent blog post, we looked at how long it takes to make one plasmid and how much time Addgene has saved ...researchers. Assuming 5 hours of hands-on time to make each plasmid, we would have saved 5 million hours...will be moving into a bigger space so that we can make an even bigger impact - we are always on the lookout...

...topics you want to learn more about. Our post on “Making Your Own Competent Cells” has been extremely popular...additional questions. We’re happy to see scientists making their own cells to save money and create the tools...Replication By Kendall Morgan Save Time and Money by Making Your Own Competent Cells By Melina Fan Plasmids...

...Neuroscience meeting in Washington, D.C., we must be making a dent in the global coffee supply… My back-of-...we've spoken with are excited and passionate about making such tools open-access. Though it is time for me..., remember there are only 2 days left for you to make the most of this great opportunity – so if you’re...

...PFA-saponin, PFA-triton). The fix/perm solutions can make a significant difference in the efficacy of your... and the small difference in protein expression makes confirmation of antibody selectivity even more challenging...used has endogenous levels of expression, this can make the results more difficult to interpret, and a knockout...Antibodies. The Only Good Antibodies community works to make best practices in research antibody selection and...

...story. This year, we made significant strides in making science more accessible and reliable. We continue...experimental results. We also redesigned our website to make it easier to find what you need and to bring it ... every breakthrough enabled brings us closer to making an impact on humanity's greatest challenges. Thank...

...is being an overthinker. Being an overthinker can make me scared of taking risks, afraid of leaping forward...in a life sciences career but life has a way of making unexpected decisions. As someone who was not necessarily...additional person each day. It felt like I could actually make connections with people. These were some of the ...

...control of an otherwise ubiquitous promoter. You could make an entirely new mouse line with mCherry controlled...paired with any Cre-controlled gene of interest makes pmSyn-BFP-Cre highly modular. Further uses of pmSyn1...other viral vector tools in the years to come. We’ll make many more viral vector tools and resources available...

...expression vector and attached a DNA barcode to each to make the library usable for pooled screens. Applications...different contexts.” It’s also possible you could make use of these plasmids in living animals, to explore... that’s thrilling to us,” Wood says. “We want to make these available as widely as possible.” The researchers...

...this interest in the ways science is shared and in making it easier to share, we take special note as the...Way to Share Science, in honor of a decade spent making plasmids more accessible and easier to share. As... As we prepare to ring in the New Year, let's make a toast to open science. And, please, share your favorite...

...capacity (8-10 kb) than other retroviral vectors. This makes it possible to use a polycistronic cassette that...Another challenge is that multiple transfection makes it difficult to control the dose of plasmid the ... pluripotency of mouse cells typically involves making chimeras by injecting iPSCs into blastocysts and... and Juan Carlos Izpisúa Belmonte. "Methods for making induced pluripotent stem cells: reprogramming à...

... the 1990s, biologists have spent a lot of time making things glow. Chances are you’ve used a GFP derivative...derivative to conduct subcellular localization studies or make a reporter construct. Fluorescent proteins (FPs)...the unexpectedly photoconvertible GFP proteins to make a new transgenic line could avoid problems in localization...

...partners and Tech Transfer Offices at universities to make COVID-19 research-related plasmids available more...Find plasmids available to industry scientists To make plasmid sharing more open for academic and industry... deposited plasmids at Addgene and would like to make them available to industry, please reach out at ...

... by making them brighter, more photostable, smaller, and less toxic to cells. They’d like to make near-infrared... muscles work inside a living animal. For those making use of these probes, he says, when in doubt, read...

...toxicity or high off target effects, both of which make it a non-ideal drug. If a drug selectively kills...added QC is where CRISPR can save time and money to make sure quality candidates are selected.      ...Additional resources on the Addgene blog CRISPR 101: Making a Knock-In Cell Line Using CRISPR-Cas9 to Edit...

...best way to keep everyone on the same page is to make communication a non-negotiable part of your collaboration...Luckily, there are techniques and tools that helped make my experience run smoothly and can help other teams...meetings can be tricky, and it takes extra effort to make sure everyone in the meeting has an equal opportunity...

...because I knew it would make me a better writer, but also because I knew it would make me a better scientist...mates, you add the individual figures and results to make the final product with an interesting introduction...

... mechanisms for feedback (more on this below). Make sure to give positive feedback when it is merited...some people find this hard so get used to it by making it a concrete goal to say “thank you, job well ...unnecessarily big issue for you and for the employee. Make sure to focus on the action, not the person - intent...

... of the features you may want to consider as you make your choice. The first and most important thing ...vector that you will need. You will also have to make sure that your plasmid has been incorporated into... applications used to study gene function. From making transgenic animals, gene targeting and genome modifications...

... of bacterial genomes utilize this system (4). Makarova et al. grouped the systems into three distinct...Let us know.  References Ishino Y, Hideo, S, Makino K, Mitsuko A, Nakata A. J Bacteriol. 1987 Dec; ...1709–1712 (2007). CRISPRdb. Date: 2013-09-18. Makarova K, Haft DH, Barrangou R, Brouns SJJ, Charpentier...

... four. No Y chromosome is present in this line, making it genetically female. HEKs are generally cultured...properties. HEK-293 cells are tumorigenic, which makes them a candidate cell line model for cancer-specific...reproducible results. It’s also widely available, which can make for a seamless transition of biological tools to...

...spatio-temporal regulation, and is difficult to reverse, making it unsuitable for some types of experiments. (To...arabinose, requires many wash steps to remove arabinose making it difficult to regulate quickly. Arabinose also...Khammash M, Di Ventura B (2021) Engineering AraC to make it responsive to light instead of arabinose. Nat...

...experiments, making AAV starts with some plasmids. You just need three plasmids to start making AAV:  the...sequence Inverted terminal repeats (ITRs) are what makes an AAV transfer plasmid an AAV transfer plasmid....

...development, vaccine development, and assay technologies makes her a perfect fit for Addgene. She was Head of Quality...around the world who would otherwise have difficulty making or getting these reagents is profound.” Frank Stegmeier...we’ve built create inefficiencies that ultimately make innovation and science slow,” Vandenberghe says....

...battery bank and the vehicle battery/alternator to make sure the vehicle battery didn't run down. In...temperatures and unstable ground around these hot springs make them extremely dangerous, we used telescoping poles...protocol. IgnaviCas9’s incredible thermostability makes the enzyme particularly useful. Additional resources...

...screening experiments. Get the full details on how to make these libraries from the post and the associated...Editing CRISPR has revolutionized how researchers make knockouts and other types of genome edits, but, ...depositors and guest writers. Your contributions make Addgene and the Addgene Blog useful resources for...