We narrowed to 871 results for: PLE

...recombination intermediates can then resolve to complete the DNA repair process Homology directed repair...happen on the crossing strand (horizontally at the purple arrows) or on the non-crossing strand (vertically...is elongated and ligated to fill the gap, thus completing SDSA.   Break-induced repair (BIR) pathway BIR... and even easy. This targeted-break technology coupled with HDR is a powerful site-specific knock-in tool... uses reverse transcriptase to generate the complementary ssDNA. Easi-CRISPR was shown to increasing editing...series was designed to help explain the basic principles driving CRISPRs and genome editing technology...Pfeiffer P, et al. Mutagenesis (2000). PubMed. Multiple Pathways of Recombination Induced by Double-Strand...

...plants using the same general experimental design principles developed in other model organisms (see our CRISPR... CRISPR guide for common CRISPR principles). However, plant-specific modifications to commonly used CRISPR...cloning required. As with other model systems, multiple gRNAs can be expressed to modify several genomic...genomic loci at once (get more information on multiplexing gRNAs here). Cas9 is commonly tagged with a nuclear...longer RNAs (such as mRNAs for gene expression). Examples of commonly used RNA pol II promoters for Cas9...delivery and expression in planta using chitosan-complexed single-walled carbon nanotube carriers. Nat Nanotechnol...Zhang Y, Qi Y (2015) A CRISPR/Cas9 Toolbox for Multiplexed Plant Genome Editing and Transcriptional Regulation...

...are designed to be complementary, but each primer may extend beyond the complementary region as long as...with enzyme “B” detects primer multimerization (purple) which is evident by an increase in the size of... Primer Design As a rule of thumb, 11 bp of complementary sequence on either side of the desired mutation...A-overhangs (a feature used in TA-cloning). Such non-complementary bases at the 3’-end interfere with plasmid reconstitution...that any residual parental plasmid (usually from incomplete DpnI digest) can also form colonies under these...particularly relevant in cases where a large plasmid with multiple functional regions is used as template for the.... Breslauer, Kenneth J., et al. "Predicting DNA duplex stability from the base sequence." Proceedings ...

...defining a couple of key terms is a better approach. Also, appealing images and colors draw people in. Drawing...webinars—I have them stacked close together for the next couple of months. This involves a lot of communication...of science communications-- Be able to talk with people: “If you do this for a living, science writing ... when I am in new territory. Perhaps there are examples of science writers who can get everything that...non-scientists: “Jargon is a killer for science literacy. If people have to understand a lot of big words to access...mechanism that also looks nice is a great way to bring people in.” Are you interested in careers outside academia...

...Optogenetics Ofer Yizhar New viral prep with multiple serotypes pAAV-Syn-FLEX-rc[ChrimsonR-tdTomato...Biosensors GENIE Project New viral prep with multiple serotypes pGP-AAV-GFAP-iGABASnFR2(no bind)...Biosensors GENIE Project New viral prep with multiple serotypes pGP-AAV-syn-iGABASnFR2-WPRE AAV1...Biosensors GENIE Project New viral prep with multiple serotypes pGP-AAV-syn-iGABASnFR2(no bind)-...Biosensors GENIE Project New viral prep with multiple serotypes pGP-AAV-syn-flex-iGABASnFR2-WPRE...Biosensors GENIE Project New viral prep with multiple serotypes Looking for more viral vectors... Multiple Authors...

... forming the repeat:antirepeat duplex. tracrRNAs also contain multiple stem loops that are recognized ...Palindromic Repeats. That contrast of simplicity and complexity is reflected in the biology, too. CRISPR is an..., and the spacer sequence is used to target complementary DNA sequences. In laboratory settings, an sgRNA...whose sequence is not checked. Assuming full complementarity, its sequence matches the guide sequence. PAM...either crRNA or sgRNA). Cas9 requires perfect complementarity in this region between the guide and target...post on the structural biology of Cas9. Want a simpler guide to keep at your desk? We’ve got a CRISPR ...

...found the people at Addgene have become my friends, mentors, and some of my favorite people to talk to...student at Northeastern University, I am required to complete a six month work experience called a co-op to ...work of others. Having an environment filled with people who are interested in understanding the work you...Thankfully, my managers set aside time for me to complete an online social media certification on social...changes to our social media strategy. We ended up implementing some of these changes and seeing early results...results that suggest our content is reaching more people who need it!  After a few months, we wanted to share...

...research project can take several years to complete and has multiple benchmarks. Think about what your personal...professional stage of your career by connecting you with people of similar interests and goals. The following questions...field? You want to choose a research question that people (especially you!) care about. Have you critically...reach these milestones? Can you get assistance from people who have the required expertise to help move your... of keeping up-to-date with the literature of a couple of well-known journals that discuss general science...Researchers should  pick a topic that “looks at the complexity of biology and break apart a piece that can address...

...not powerful enough for large brain samples. For large brain samples, we recommend pretreatment with ScaleCUBIC1...fluorescent protein (XFP) genes in a transgene. When multiple copies of the transgene cassette are introduced...resulting in low fluorescent protein level. The principle of Tetbow In modern neuroscience, plasmid or viral...time points, sacrifice the animals and fix the samples. When you use AAVs, it typically takes a few weeks...excessive amount of XFPs expressed. Then, clear the samples and visualize. For more details on this procedure..."Optical clearing and index matching of tissue samples for High-resolution fluorescence imaging using ...

...transparently reveal them upon completing an experiment. There’s not necessarily in-principle acceptance, but readers...simply doing an experiment wrong. However, even simple procedures like pouring a plate or running a gel...sifting through paper lab notebooks). Once you’ve completed your experiment, you’ll need to analyze your results...individuals and by organizations like Addgene (a good example is our series of blog posts on the defunct genome...promote reproducible publishing practices. One example is the publication of peer reviews so we can see...and, if approved, the researcher would get in-principle acceptance of the final manuscript regardless ...interlab peer review of experimental design. These are simple steps but could go a long way to encouraging researchers...

...where we aim to do large scale DNA preps with multiple samples without cross contamination. We currently ... in addition to preparing DNA and generating/replenishing kits, is to find new ways to make lab processes...futuristic with block chairs in the hallway for people to take a break and relax. Companies at LabCentral...create a DNA library, one must first fragment a DNA sample into many small components. Without proper processing...process with, we got great amplification of our sample libraries. NEB also gave us a few goodies which... I am bombarded with a lot of information. For example, before going to my next workshop, I’ll ask myself...

...Controlling multiple genes with Gal4/UAS system One can also use Gal4 to drive expression of multiple genes ...the Gal4/UAS system. This incredibly useful, yet simple system allows you to systematically study gene ...downstream of a tissue-specific promoter sounds simpler, this modular system saves you from having to create... making a fly line in the next blog post). For example, the hedgehog gene can be overexpressed in several...under the control of UAS enhancer sequences. For example, if you discovered five new genes and wanted to...control purposes that there are dosage effects. For example, overexpression of two genes by one Gal4 driver...

...disease affects multiple organ systems. Nevertheless, the findings are proof-of-principle for use of CRISPR... bulk of which you can find below. CRISPR for complex genetic diseases As CRISPR technology becomes increasingly...using it to investigate potential therapeutics for complex genetic diseases. So we’re revisiting this post...look at CRISPR’s potential for use in treating a complex genetic diseases, Alzhiemer’s disease (AD).  Troy...diseases with real phenotypic effect.” “It’s a very simple technology,” Doudna further explained. “That’s ...  Multiple Authors...

...the requesters what they needed. In the example above, for example, we fostered a formal scientific collaboration...uploaded to the preprint repository BioRxiv. This example clearly illustrates the power of collaboration,...choice but to quarantine in our houses, feeling helpless about the situation. But a few days later we had...certain projects. The idea behind the platform was simple: any person working on COVID-19 could file a request...groups. In other cases, the collaborations were simpler and the volunteers helped with things like translations...need help with your scientific research. It’s completely free, of course. You can find more information...

...while working remotely. Social distancing? Many people are now calling this “physical distancing” since... important to stay virtually connected with the people in your life. At Addgene, we’ve started virtual...SocialDistancing may send a wrong message for some people. We should recommend #PhysicalDistancing. Social...take-out) or volunteering to foster a pet, for example. If you are looking for some resources specific...activities that kids can do more independently. For example, they could do something like puzzling or crafts... time.  Find learning activities for kids. For example, Skype a Scientist is now open to families at home... future experiments and other researchers by implementing reproducibility best practices in your research...

...realized that it was easy to find people who were interested. For example, our journal club discussion on... CRISPR/Cas9 gene drive paper was very popular. People kept asking if we were teaching classes at the ... safety training prior to getting started. For example, Timothy Stiles joined BosLab last year with a ... space to perform early-stage experiments. For example, Dave Grawoig is using nanopore sequencing at BosLab... getting the community to collect various soil samples, extract DNA, sequence it, and analyze the data... characterize the microbiomes of the different samples. The goal is to not only learn something about ...

...understanding these complex processes can’t be overstated, we can also learn a lot from Nature’s simpler solutions...engineer it to suit our purposes? In search of simple biological machines R bodies can repeatedly ...These questions can be addressed using R bodies: simple, force-generating protein machines that can extend...could be used in a variety of applications. For example, you could use them to lyse cells (and surrounding...

...time for that and you won’t be as productive. For example, the many resources available on the Addgene website...less dependent on the manager to succeed. More people in leadership roles leads to more ideas, initiative...enough to do…you are not delegating enough. For example, if we need to develop a new plasmid collection...duplicated. With each component of the project completed by an individual who is a local expert in that...result is high quality and the entire process is completed more efficiently. Listen to our podcast interview... Don’t give out tasks – where possible, include people in deciding what is to be delegated to them. Not...your time and skills. It also ensures that other people in the team grow and develop to reach their full...

...Chemical agents, temperature, and light are all examples of factors that can lead to the induction of a... these three types of inducible promoters, and examples of each type. Many of these promoter systems are...transcription to begin. In addition to arabinose, cAMP complexed with cAMP activator protein (CAP) can also stimulate...stimulate AraC binding to I1 and I2 sites. Supplementing cell growth media with glucose decreases cAMP ...represses pBad, decreasing promoter leakiness.   Other examples of chemically induced promoters include positive...in plant research. Promoter Subtype Promoter Example Activator Addgene Plasmid/Kit Alcohol-regulated...temperatures but can be induced by heat or cold exposure. Examples include the heat shock-inducible Hsp70 or Hsp90...

... to determine precisely how the methods were implemented. We run into this problem frequently over the...not robust. Sampling noise There is, of course, simple noise in the process of sampling and inference-...problems above: unclear methods, QRPs, and small samples, then when replications do not come out the same...statistical tools that generate a p-value.  As an example, if you hypothesize that growth rates will be higher...answer those questions, the article is given “in-principle acceptance” to publish the final results regardless... special emphasis on positive controls and good sample sizes, concerns that are too easy to overlook when... worked with citizen scientists to design and implement authentic scientific research and as the Director...