...you’ve been able to check out some of the many applications of AAV in some of our other blog posts and ...AAVs) make fantastic gene delivery vehicles for episomal gene expression and are particularly useful for...editing system frequently over the past fifteen years. Some examples from their lab include using AAV to introduce...and have also been able to browse some of the ready to use AAV available through our viral service. In addition...Manu, et al. "Facile methods for generating human somatic cell gene knockouts using recombinant adeno‐associated...Jung-Sik, et al. "Intact cohesion, anaphase, and chromosome segregation in human cells harboring tumor-derived...engineering using adeno-associated virus (AAV)." Chromosomal Mutagenesis. Humana Press, New York, NY, 2015...

... to give the audience something to think about, argue with, or learn from. Some of the best talks I have.... Two of my jobs have indirectly resulted from someone seeing me speak in a non-interview setting. There... resources on self-promotion (how hard it is for some people, especially women), visibility (how to get...comes. 5. Choose interesting content Talk about something you know a lot about and that you care about. ...company you are visiting works on them. Of course, if some of your work does relate, make sure to include that...with your confidence. It is best not to be asked something that you don't know the answer to, but if you ...speakers don’t have to work at it. You have to put in some effort to be the best speaker you can be. Seek out...

...protein domains to the appropriate locus has been cumbersome. To overcome many of the limitations intrinsic...regulation has roles in many cellular functions, from endosomal trafficking to signaling pathways to the cell ...codons from the antibiotic resistance cassette, a ribosome binding site, SacI restriction site and a linker...system utilizes the bulldozer-like quality of the ribosome during translation, which knocks off all mRNA ...mammalian cells. Multicolor CRISPR labeling chromosomal loci in genome Thoru Pederson’s lab at the University...dynamic interactions between intra- and inter-chromosomal domains. The authors anticipate that this tool..., genome rearrangements and cancer-associated chromosome shattering. This method is based on the recognition...

... parts of CRISPR. The bacterial chromosome encodes a tracrRNA (in some systems including Cas9), Cas proteins...DNA, whereas some mismatches can be tolerated outside this region. Conclusion Ready for some more details...DNA CRISPR array: The section of the bacterial chromosome that stores a genetic record of previously encountered...by the Cas9 protein from the appropriate species. Some Cas proteins, like LbCas12a or AsCas12a, do not ...system for laboratory applications. At Addgene, we sometimes refer to the programmable guide sequence as the...

...building strong connections with coworkers and doing something meaningful with my career. I can’t pinpoint...summer travel plans while eating lunch. There was something so special about her pure interest and curiosity...biweekly report of our social media analytics, something that had never been done at Addgene before. This... social media strategy. We ended up implementing some of these changes and seeing early results that suggest... and trampoline all at once. I was able to do something that was challenging while helping us share our... at Addgene have become my friends, mentors, and some of my favorite people to talk to. This six month...confident in myself, my abilities, and my future - something I didn’t expect from a co-op experience.  Sarah...

...better with some variety in their job. Good relationships with clients and colleagues Sometimes this means...skills. Managing a team is hard and most people have some learning they need to do before they can be good...type of bad manager have you had?). You can avoid some of these common pitfalls of management and still...when making decisions, assignments and polices. Sometimes the boss has to be the boss and make difficult...because, let’s face it, we are special. Below are some special characteristics of scientists in the work...deadlines (there’s that need for flexibility, sometimes taken to an extreme by scientists) Are highly ...

...recommendations on some ways to get started sharing your science with the community. Some of them you can...ask and answer questions and hopefully discover something new. Science involves success and, more often,...not just the details of your science but also something about how science really works. 2)   Read science...your own twitter feed or talking about and sharing some science on the one you already have. Read my earlier...SciFund Challenge. While the goal is ostensibly to get some funding for your research, SciFund is really motivated...finally understood why he cared to study mice on some distant island. He got a little money for his trouble...

...2017). With the right conditions and parameters, some people find it can detect even smaller differences...copy of a transcript. But this sensitivity can sometimes cause as many issues as it solves, so understanding... could easily be its own deep dive, but here are some general helpful principles (and I recommend Bustin... multiple reference genes whenever possible (Vandesompele, 2002), and it's possible to multiplex your ...documented in the literature, a geNorm study (Vandesompele, 2002) will help you select the appropriate ...produce reliable and reproducible data. Here are some tips to help you in your execution:  Accuracy is...Passive Reference Dyes Whatever your pipetting method, some variation will occur. A passive reference dye, such...

...serotypes have a primary cellular receptor and sometimes one or multiple coreceptors, which may be present...present on different cell types and sometimes vary between target species or even strains. Pro tip! Getting...serotype for Addgene’s Packaged on Request service. Some AAV1 tools are available through our in-stock viral...serotype for Addgene’s Packaged on Request service. Some AAV2 tools are available through our in-stock viral...serotype for Addgene’s Packaged on Request service. Some AAV5 tools are available through our in-stock viral...serotype for Addgene’s Packaged on Request service. Some AAV8 tools are available through our in-stock viral...available through our Packaged on Request service, but some AAV9 tools are available as in-stock preparations...

...support your favorite local exercise spot or find something new. We usually have yoga class twice a week at... #SocialDistancing may send a wrong message for some people. We should recommend #PhysicalDistancing....those of you who have the desire and ability to, some communities are looking for bench scientists to ...foster a pet, for example. If you are looking for some resources specific to self-care during the COVID...networking you can do online. See if a scientist or someone you’re connected with would like to do an informational...more independently. For example, they could do something like puzzling or crafts, while listening to podcasts...productive as you would be in a normal situation. For some other resources on working from home, check out ...

...Relationships need nurturing. Perhaps you’ve met someone interesting at an event. Or, perhaps you have a...ways to strengthen your new relationships Here are some ways to go about strengthening a new relationship...there’s good news.  Send an email if you have something to ask or say. It can be as simple as “it was ...appreciation. Send a thank you note or gift to someone who helped you succeed in any way. Don’t believe...strawberries for helping with an introduction that helped someone get a job interview. Even just a thank you email...is having a mutual acquaintance connect you to someone who you need to speak to for informational or career...ask me about the different arms of this starfish sometime, but as of yesterday, I have 4,700 LinkedIn connections...

...sites on different chromosomes in living cells, the system requires high-copy chromosome-specific loci in...study the spatial and temporal arrangement of chromosomes and how these arrangements may affect nuclear...based multicolor labeling system to visualize chromosomal loci in human cells (Ma et al., 2015). To do ...inter-locus distances. They tested this system out on chromosome 19 by targeting 46 repetitive loci that contained... (PR1), the p-arm (T1), and the q-arm (T2) of chromosome 19. Image from Ma et al., 2018 with permission...Hanhui et al. “Multicolor CRISPR labeling of chromosomal loci in human cells.” Proceedings of the National...

... allows it to replicate episomally, essentially remaining an extrachromosomal part of the host genome ...single-stranded DNA parvovirus. AAV can infect humans and some other mammals but is not currently known to cause...This is large enough for many applications, but in some cases multiple AAVs (co-infection), an alternative...humans, many mammals, and even the avian population. Some adenoviruses are known to cause respiratory disease...Adenovirus generally has high immunogenicity, but some strains have been engineered to induce a lower inflammatory...AAV’s several years). But these viruses are roomy: some adenoviruses pack up to ~35 kb of space for cargo...

... next steps in the process involve repairing chromosomal DSBs. It is important to know that cells possess...subunit DNA ligase (XRCC4-ligase IV); together with some accessory factors (PAXX, XLF), this complex holds...two-thirds chance of causing a frameshift mutation. Of some importance, the deletion can be less heterogeneous...should possess indels in the majority of their chromosomes within a day. Another factor expected to impact...or more can simultaneously introduce a pair of chromosome breaks, and could result in deletion of the intervening...Stark JM (2009) Limiting the Persistence of a Chromosome Break Diminishes Its Mutagenic Potential. PLoS...EA, Giovannangeli C, Jasin M, Brunet E (2014) Chromosomal Translocations in Human Cells Are Generated by...

...also has the flexibility to modify the E. coli chromosome, plasmid DNA or BAC DNA.  To use the lambda ...linked strains contain the Lambda Red system).  In some of these strains, expression of exo, beta and gam....   This setup can be also be used to modify chromosomal genes. After the initial editing event, the defective... defective prophage can be removed from the chromosome of the host E. coli by a second lambda red recombination... system is best suited for editing bacterial chromosomal DNA because it is easy to remove the recombineering...responsible for excision of mini-λ from the host’s chromosome. After this, mini-λ can readily be purified from... being targeted for modification: the E. coli chromosome, a plasmid, or a BAC.    Pro Tip: ssDNA substrates...

.... While HaloTag reacts faster for some dyes, others, especially some nonfluorescent ligands, react more... protein, a fancy new green fluorescent protein, some affinity purification tags, another version with...with that same red fluorescent protein somebody made because they forgot it was already in the freezer… ... Common self-labeling tags Let’s take a look at some popular self-labeling tools: Tetracysteine: The ... Easy CLIP-tag 182 Benzylcytosine (BC) Some Easy Ligases Small, varies Various Few...the cells make a fluorescent protein for you. And some ligands just aren’t going to work in certain applications...check that your fusion protein is functional. In some cases, a small peptide tag like tetracysteine might...

.... These are just some of the most common parametric comparison tests along with some general statistics...long-awaited data – and it looks great! There are some exciting trends in your experimental groups, but...these designations all refer to the same test. On some occasions, there is an experimental group being ...However, it will only tell you if differences exist somewhere within your data; it does not indicate which groups...Post hoc tests Knowing that significance exists somewhere amongst multiple groups is rarely enough information...every pairwise comparison within your population? Sometimes this can’t be avoided, but if it can be, you can...working with, graphing data with a histogram is sometimes sufficient to determine normality (or lack thereof...

...universities' decisions to alter postdoc salaries - in some cases reversing these decisions entirely. At Future...are indeed going ahead with their planned raises, some are taking the opportunity of the lapsed mandate...being compelled at the state level, in the case of some public institutions) to cancel plans to raise salaries... in light of the injunction. In addition, PIs at some of the institutions cancelling plans are going ahead...communication of their plans (many postdocs - and in some cases, faculty - have found out about what their...they have in their salaries and benefits, and how some institutions are paying lip service to improving... not raising salaries when all others in Boston (some of them literally across the street from these institutions...

...relationship. Pro tip: Once you are connected with someone, their email address is usually visible to you ...if you don’t know clearly why you are inviting someone to connect, perhaps you should be asking yourself...discussions.  Pro tip: Instead of connecting with someone you don’t really know, follow instead of connecting...giving you a way to connect with them in the future. Some profiles have a blue Follow button. For less famous...do an easy favor for people in your network. If someone has connected with me, I consider them fair game...updating it. LinkedIn may actually replace the resume some day (I’ve already abandoned my paper resume). It...looking for a new job! I have been able to tell when some start-ups were about to be bought by seeing certain...

...more than one protein in your sample? Or what if something weird happened during your western and your results... targets you plan to probe for. Even using PVDF, some of your target protein will be removed from the ... first to preserve your sample. Pro tip! Because some protein is lost each time you strip a blot, you ...experiment. Remember, stripping your membrane will remove some of your sample —and harsher stripping means more... need longer incubation for effective stripping. Some blots benefit from an additional 5–10 minutes of...blotting that you’re looking for a “blank” result! If some signal remains, you may want to repeat your stripping...strip and reprobe your western blot several times — some researchers report up to ten cycles! Just remember...