... 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 ... can skip the first round of production, saving about a week of work. An important note is that this is...

...our tissues unimpeded. Andrew and Maria dreamed about the possibilities of a magnetoresponsive fluorescent...of improved response. Then, just when Andrew was about to suggest she deprioritize the project, Maria’s...with a strong commitment to exploration and collaboration. Andrew is a physicist with a background in ...potential applications.  Andrew is a frequent collaborator and is happy to give advice and support for ...interested in working on these questions or collaborating, the York Lab is eager to hear from you. Find...

...all the current code on GitHub, ask us questions about it, or send us a pull request for improving it. ...) so that they can be used in the rank equation above. Cut-to-insert distance was fitted to experimental...below) so that it can be used in the rank equation above. Generally, the greater this distance between the...splits up the guide sequence.) If neither of the above are options, the algorithm silently mutates codons...Addgene blog Read our CRISPR 101 blog posts Learn about some considerations when designing gRNAs Find other...

...positive experience, I was still incredibly nervous about leaving academia. A lot of academics tend to view...which may take a longer time to acquire. Learning about drug discovery: Unless your academic lab is closely...information. Working as a team: Industry is all about teamwork. A single individual could not bring a ...someone with whom you can have an in-depth discussion about your project might be challenging. In an academic...contacts to get that referral and find out more about the company’s program. Once you’ve found a few positions...

... think much about management skills during your scientific training. I never thought about management ...open mind. First let’s clear up a misconception about being a manager: yes, being a manager means being...you should be doing : writing a grant, thinking about the next project, reaching out to other teams, expanding...buffer them from unnecessary pressure “coming from above”, which is when other skills such as negotiation...

...Until then, you can only wonder what I am talking about. How do researchers use physical models of protein...students work closely with local researchers to learn about their research while creating physical models of...model is an effective teaching tool. You can read about Jacob’s initial reaction to this model on his blog...will wake up in the middle of the night dreaming about the conformational gymnastics that this protein ...Additional Resources on the Addgene Blog Learn All about CRISPR Use CRISPR for Genome Visualization Tips ...

...If you follow CRISPR research, you know all about using non-homologous end-joining (NHEJ) to make deletions...plasmids, GFP knock-in cell lines can be created in about a month and a half, without the need for complicated...more predictable than NHEJ. As seen in the figure above, short (5-25 bp) regions of homology flanking a ...Directed Repair and Non-homologous End Joining Read About Other Genome Engineering Tools like Sleeping Beauty... Additional Resources on Addgene.org Learn All about CRISPR on Our CRISPR Guide Pages Find CRISPR Plasmids...

...republished here with permission. Quilt is a collaborative database for genomics. In this article, Quilt...tri_methylation In the next section we’ll apply the above formula to a real-world experiment. We’ll start ...discovery and therapeutics. We’ll have more to say about clinical applications of CRISPR in the next section...Aneesh Karve is CTO at Quilt Data. Quilt is a collaborative database. Aneesh's research interests span machine...on Your CRISPR Background with CRISPR 101 Learn about CRISPR Delivery Methods Validate Your Genome Edit...

...a less tangible form: knowledge and information about the materials in our repository. We started to increase...by making it easier to find materials and learn about them. As early as 2007, we began grouping similar...professors, stopping by to say, “Everything I know about virus, I learned from Addgene,” or “I learned to...seeing our resources shared via social media, on collaborators’ websites, or as a recommendation in a Reddit...updating and expanding our website, we’ve learned about the importance of smaller, systemic changes, like...

...a single grant. Either way, think realistically about the time and resources required to feasibly accomplish...interrupted frequently by committee members. As mentioned above, plan to give your 10 minute overview and address... list of common questions, check out this eBook about applying for faculty positions. You need to practice...program. You should be proud of it and be excited about getting the opportunity to realize your vision. ...blog posts Resources on Addgene.org Learn more about careers at Addgene Watch our career videos ...

...gene, Xw, except for a single base. If you heard about CRISPR, one question probably comes to mind: can...compatibility All of the base editors mentioned above are SpCas9 nickase-based and depend on the presence... HDR methods for rice crop improvement in the laboratory of Yinong Yang at PSU.   References 1. Komor,...The CRISPR-Cas Revolution, Cold Spring Harbor Laboratory (2018). Additional resources on the Addgene blog...Download the CRISPR 101 eBook Read our blog posts about plant biology Resources on Addgene.org Find all...

...brainstorming sessions.  Allow the group time to think about the problem before you meet. This holds the dual... meetings run smoother when people have thought about the issue ahead of time. Encourage dissent. The ... decisions while encouraging the group to think about alternative options. When a devil’s advocate is ...a few minutes every month dedicated to thinking about the future will improve your ability to innovate...to be able to continue our experiments. Thinking about these plans before you’ve run out of consumables...

...101 blog posts Learn more about the origin of replication Find blog posts about plasmid cloning Resources... plasmids within the same bacterial cell, think about whether plasmid incompatibility will affect your...this is expanding as we obtain more genomic data about plasmids (Shintani et al., 2015). Now, bioinformatics...of the microbiological world. (Want to know more about these plasmid types? Read our Plasmids 101: Environmental...

...target metabolite creates an aptamer that will only bind to DFHBI in the presence of that metabolite (Fig...approach less sensitive in most applications. Learn About Fluorescent Biosensors Light-up aptamers (or, fluorescing...fluorophore protein. Years later, Samie Jaffrey’s laboratory took advantage of their deep knowledge of the...Tips for Using FRET in Your Experiments Learn more about GFP Controlling Protein Activity with GFP Resources...

...complete editing process. The MUCICAT system While the above systems are great for experiments that require only...serve to interrupt genes (or gene clusters) in metabolic engineering. So far, we’ve used the MUCICAT system...when compared to the industrial strain. In our metabolic engineering case study of N-acetylglucosamine ...Multiple induction systems to choose from. Ready for metabolic engineering with improved cargo dedicated to gene...other CRISPR methods for bacteria Find all articles about microbiology Resources on Addgene.org: Visit the...

...deletions, the ability to analyze and interpret data about these types of edits is critical to the CRISPR workflow...Distribution and Traces - provides particular details about the indel profile of the edited sample. You can ...sequence base calls from the .ab1 file are also shown above each trace. The horizontal black underlined region...us know how it works for you! Read more details about the ICE tool in our bioRxiv paper. The new ICE CRISPR...guiding CRISPR researchers, she's either day-dreaming about exploring sci-fi worlds or exploring nature in the...

...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 ... times and on different days. Using our example above, we would test different preparations of Plasmid...

...time I heard about FRET during a journal club, my guitarist brain automatically thought about the raised...you would say. The student was of course talking about the now well-known FRET, aka Fluorescence (Förster...other. FRET reduced the scale of our perception about molecular interactions. What is FRET? In their ...Resonance Energy Transfer    Interesting Articles About FRET: Piston DW, Kremers GJ. Fluorescent protein...

...Homology-Directed Repair Learn about base editing for plants Read about four base editor reporters Additional... The BE3 system increased editing frequency to above 30% for a variety of targets in human cell lines... Liu lab created BE4max and AncBE4max mentioned above, they also made an adenine base editor with improved...cell therapy applications. Dual base editors What about combining the function of both editors into one?...also RNA base editors, which you can learn more about in this blog post: CRISPR 101: RNA Editing with ...

... this chain (blue) to infer or calculate things about earlier ones. In practice, multiple sources of error...condition 2.” By eye, the loading control appears about the same between the two conditions. But it isn’.... However, it throws both assumptions discussed above into question. Assumption #1: The loading control...searching the literature to see what is already known about your housekeeping protein in your context. If you...step from the POI blot, you don’t need to worry about the linear detection ranges of multiple targets....