...about a month and a half, without the need for complicated cloning of homology arms. MMEJ: An Introduction... inserted into the PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an insertion locus-specific... for making targeted modifications. A recent publication by Zhang et al. shows just that - using MMEJ ...lies in the diversity of nucleases and their applications. It will be interesting to see what new editing...PMID: 26678082. Find the plasmids from this publication at Addgene. 2. Dalvai, Mathieu, et al. (2015...633. PMID: 26456817. Find plasmids from this publication at Addgene. 3. Zhang, Chi, Xiuhua Meng, Xiaolei...

...delivered with an enhanced pU6cm promoter, viral replicon amplification of cargo, and heat treatment.  ...2024 bioRxiv, under CC-BY-NC-ND 4.0 International license. With this approach, the team was able to drive...Arabidopsis, suggesting the improvements may be broadly applicable to dicot species. In addition, they generated...Adapted from Smith et al. 2024 under a CC-BY 4.0 license.  The team deposited an array of constructs...activation triggered by endogenous GPCR. Example applications include large-scale pharmacological profiling...adapted from Wu and Wang 2024 under a CC-BY 4.0 license. Find more neuroscience antibodies here! ...

... primarily on fluorescent proteins and their applications, tackles much broader issues in this brief volume...sexism in science, problems with peer-reviewed publications and funding, bioethics in the age of CRISPR,...to the steadily increasing distrust the general public has in science and scientists (as discussed in ...Readers unfamiliar with academia’s challenges with publication and funding may be dismayed to see the dirty ...CRISPR of course address some of the ethical implications of using these technologies, but also the rapidity...and the marginalization of science, particularly public health and environmental science, by the Trump ...

...emerging work by collaboratively discussing recent publications in a specific field(s). Most anyone who has ...major findings and then focusing more on broader implications rather than diving deep into the individual ....  These variations are suited for different applications and should be carefully chosen for each group...first meeting, now what? After the first meeting solicit feedback. To ensure that the journal club is useful...wanted, but were unable to attend for scheduling conflicts. This helped us find a time that accommodates ...accommodates more people. The feedback could be solicited via multiple avenues including casual request via email...

...production system may depend on what downstream applications you are planning, cost, and the amount of protein...accurate concentration to give your downstream applications the best chance at being successful.  Using ...they bind their antigens specifically. Possible applications for validation include western blot, immunohistochemistry...some antibodies are better suited for certain applications. An antibody that performs well in immunohistochemistry...use! You can learn more about common antibody applications in our Antibodies 101 blog series.  Resources...H, Zhang Y, Zhan C, Malm M, Field R, Turner R, Sellick C, Varley P, Rockberg J, Chotteau V. Small-scale...

...-target editing in a cell culture model. The application of Cpf1 to genome editing is exciting both in...in terms of basic science and translational applications. This discovery of this type V CRISPR system ...PubMed PMID: 26422227. Find plasmids from this publication at Addgene. 2. Zetsche, Bernd, et al. "Multiplex...PubMed PMID: 27918548. Find plasmids from this publication at Addgene. 3. Kim, Daesik, et al. Genome-wide...PubMed PMID: 27272384 Find plasmids from this publication at Addgene. Kleinstiver, Benjamin P., et al....PubMed PMID: 27347757. Find plasmids from this publication at Addgene. Makarova, Kira S., et al. "An updated...

... Aptamer biosensors are used in a variety of applications, including detection of environmental pollutants... opens aptamers for use in many entirely new applications including aptamer based fluorescent biosensors... render this approach less sensitive in most applications. Learn About Fluorescent Biosensors Light-up..., MG proved to be too toxic for many in vivo applications, but this work proved the concept that RNA could...and biochips. Ultimately, there are just some applications that lend themselves to aptamer-based instead...Kyung-Mi, et al. “Aptamers and Their Biological Applications.” Sensors (Basel). 2012; 12(1): 612–631. PubMed...

...each sample by clicking on the sample name or on its corresponding bar graph entry. Clicking to initiate ...sequencing data alongside the analysis of NGS-based amplicon sequencing data and found that the accuracy of...100-fold reduction in cost relative to NGS-based amplicon sequencing. To use the tool, you start by uploading...are no parameters that need optimizing and no complicated steps to learn. For increased flexibility and... download the entire analysis as a ZIP file by clicking the Download Analysis Data button on the bottom...

...use, functional in most species, and has many applications. That said, SpCas9 is not the only game in town... makes this molecule unique, and the various applications of Cas13a. The origins of Cas13a: An RNA cleaving...discussed below. Subscribe to CRISPR Blog Posts Applications of Cas13a   Figure 1: Using Cas13a as...of nucleotides.   What are the potential applications of Cas13a given what we know about its structure...fusion proteins for imaging, tracking, modulating splicing, and regulating expression of specifically targeted...CRISPR toolbox and will continue to expand the applications of CRISPR. Since this post was originally published...

...investments over the next 3, 5, or 10 years. Complicating these challenges is the fact that they occur...chemotherapies, or developing innovative mobile phone applications and services that help diabetic patients manage...collaborative work place, challenges can arise due to conflicting work styles, team situations, and personalities...literature, and critical analyses skills were applicable to my cases. I can directly use my understanding...the gap between the science and the commercial applications of a particular therapy. With your scientific...conversations, reading online healthcare industry publications like FiercePharma, FierceBiotech, and BioCentury...

...strategies Publication System nickname Photoactivatable moiety Cas9 variatn CRISPR protein application Split...improvements to CRISPR technology since its initial applications for genome engineering in 2012.  Many of these...particular interest since blue light has limited applications in vivo. UV and blue light is readily absorbed..., allowing the release of Cas9. A noteworthy application of the CASANOVA system was its use in light-mediated...continue to evolve and can’t wait to see what cool applications you use these for in the future! References...Bouazza B, Schumacher CH, Bratovič M, Charpentier E, Möglich A (2013) Engineering of temperature- and light-...

...include understanding of allelic variation between different plant species and metabolic properties affecting... ease of manipulation and their ability to be replicated. Because E. coli are easily propagated and have...minutes, they are perfect for molecular cloning and metabolic engineering.  The E. coli genome has an extensive...Escherichia coli as a Model Organism and Its Application in Biotechnology. In Escherichia coli - Recent...Physiology, Pathogenesis and Biotechnological Applications. InTech. https://doi.org/10.5772/67306 Koornneef...

...molecular biology applications for biotin and avidin. A short list of some other common applications and links...downstream application. Biotin labelled oligonucleotides can be used in a multitude of applications including...covalently attached to the active site of key metabolic enzymes including biotin carboxylase and decarboxylases... feng, Liu, N. hua, Shu, L. yuan, & Sun, H. “Application of avidin-biotin technology to improve cell adhesion...

...data, choosing controls, or mixing clear solutions—public speaking is skill that any scientist can learn....room, so it is a good idea to start thinking about public speaking early and often. Two of my jobs have indirectly... and note how they build suspense. Notice the simplicity of their slides and how the slides complement...of useful, entertaining science lab, career, and policy content. Watch Joanne give her "Not" Networking... done so. How can you expect to deliver a great public presentation if you are doing it for the first ...about how the sharing model works. See if you can elicit helpful comments from your audience as in “Does...

...reservoirs, chronic inflammation, and negative metabolic effects of ART. Why can’t the body mount an immune...create mutations in genes essential to HIV-1 replication. However, this approach could permit some viral...lessons learned in pursuing one technological application can benefit many others and accelerate the pace...: PMC4633726 Find SAM plasmids used in this publication at Addgene. Kaminski R, Chen Y, Fischer T, Tedaldi...and lentiviral packaging plasmids used in this publication at Addgene. Wang Z, Pan Q, Gendron P, Zhu W,...CRISPR/Cas9-Derived Mutations Both Inhibit HIV-1 Replication and Accelerate Viral Escape. Cell Rep. 2016 Apr...

..., the preferred vector, contains a promoterless splice acceptor-IRES-neoR gene, which enhances the efficiency...with a homologous repair template. It is this application - the creation of sequence-specific knockins ... allele of a single gene at once. Therefore, applications that require simultaneous modification of all...clones, making it unsuitable for most in vivo applications.  Third, even in the most optimized tissue-culture...efficiently deliver large repair templates for applications that require large gene fusions. The current...you’ve been able to check out some of the many applications of AAV in some of our other blog posts and have...

... do need multiple first/corresponding author publications in quality journals. It helps to come from the...a job. The currency of scientific success is publications and grants. 2. Write succinct and easy to understand...Attendees will include grad students (that are solicited for feedback), possibly the Dean, and even some...essential for human health. Failure to do so is implicated in several human diseases. Proper patterning ...disease treatment, etc., again in clear terms. "Implications for human disease" is vague. Instead, consider...interviews. For point of reference, I submitted my applications all by snail mail and only had phone interviews...

...exclude all duplicate events (two cells stuck together) from your single cells, as duplicates can’t be reliably...identify cells, the single cell gate to exclude duplicate events, and the Live/Dead gate to focus your analysis...cells, my marker stains of interest). Let’s double-click inside our gate and proceed to the next plot! Pro...show up in a straight line one the plot, while duplicates deviate from that pattern. A.                                                                                                         ... use H vs. A to distinguish single cells from duplicates. Lastly, you learned which gates should always...

...DNA-binding domains" publication.  If you want to see this new feature in action, click here to explore the...'s Addgene ID # has been referenced in other publications, you'll find a link to the list of citing articles... what I mean. Additional features Once you've clicked on the "# References" link under the "Resource ... citing articles. From the article list you can click on the PubMed link to find the article abstract ...

...search, first pay attention to the validated applications listed on the vendor’s website and choose an...western and vice versa.  In addition to validated applications, pay close attention to the controls listed ...and incubation times. If the vendor includes publications that cited the antibody, consider reviewing ... the materials and methods sections of those publications.  Finally, always verify that the chosen antibody...choose antibodies that are validated for your application and test the antibody in your specific system...to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences...

...performed on one register at a time. If a particular application makes it difficult to isolate single registers...state read-out method may not be suitable for applications that require minimal cellular perturbation. ...The input plasmid (high copy, ColE1 origin of replication), contains large serine recombinase genes downstream...contains the register. Depending on the desired application, the inducible promoters we used to drive recombinase...program to find the one that best suits your application. How efficiently do RSMs work? We tested a two-input...anticipate that, depending on the context and application, some inducible recombinase systems may work ...

...Figure 1. Used under Creative Commons Attribution License. Within the nervous system, remember that some...Creative Commons Attribution 4.0 International License.   Inducible promoters  This type of promoter.... A. (2023). Various AAV Serotypes and Their Applications in Gene Therapy: An Overview. Cells, 12(5), ...Anterograde Transsynaptic Tagging: Mapping Corticocollicular Input-Defined Neural Pathways for Defense ...Retrograde Serotype Viral Vectors 101: Viral Applications...

...additional origins of replication compared to monomers, they are also able to replicate at a higher frequency...the right, with the entire monomeric sequence duplicated in tandem, resulting in a plasmid that is twice...Characterization of multiple circular DNA forms of colicinogenic factor E-1 from Proteus mirabilis. Biochemistry...plasmid DNA production platform incorporating low metabolic burden seed-stock and fed-batch fermentation processes...What is a plasmid? Plasmids 101: Origin of ReplicationPlasmids 101: PCR...

... plasmid or group of plasmids' functions and applications. We hope that these articles make it easier ...FLIM (Fluorescence Lifetime Imaging Microscopy) applications. To demonstrate the power of mCyFP1, the Lin... Coselection efficiency is so high that some applications may permit working with edited “pools” of cells...be found at Addgene and in Figure S12 of the publication. Agudelo D, et al. Nature Methods. 2017. PubMed...gels. They are shared through Addgene without licensing restrictions. Use these instructions to prepare... subtilis has a fully sequenced genome, many applications in biotech, and is considered the most studied...regulation to genome engineering that enables new applications, such as reversing the effects of dCas9 binding...

...Addgene ID number, your publication will be listed on our website as a “Publication Citing this Plasmid” ...upon other researchers being able to find their publications in the first place. As a curator of the thousands...make it easier for you to find the plasmids and publications relevant to your research interests. We do this...number as well as the reference for its original publication. To help facilitate this process, we explain ...

... naked and unmodified dsRNA elicits immunogenic reactions that complicate the effectiveness and interpretation...silencing can be easily induced by exogenous application of long or short dsRNA. However, it is important...have benefited from the ease of use in dsRNA applications. Figure 1: Schematic of an RNAi-inducing...Historically, gene silencing methods were thought to be afflicted by pervasive off-target effects. This conclusion...LIIbeta F 1-2 RNAi, which allows assembly of intron-spliced hpRNAs through Golden Gate cloning. Artificial ...

...of the applicability and importance of research in basic biology, medicine, epidemiology, public health...the experience would improve my scientific and application writing skills, and ended up with the award. ...fantastic for broadening your experiences! These applications can be highly competitive, so asking for help...these conferences to answer questions and offer application fee waivers. One of the blog posts that helped...dedicated to different career options from science policy to science communication and much more. I hope ...

...tag to remain on your protein for downstream applications Below, we describe a few of the most common...not to perturb protein function for downstream applications. Tag cleavage must be performed after primary...also provides a visual detection mechanism via applications such as western blot and immunofluorescence....dimerizes in solution, which can affect downstream applications. Also, purifying GST by affinity chromatography... are generally best suited for their primary application. Ready to start tagging? We are here to help ...

...worms, zebrafish, frogs, some bacteria; but not thermophilic (high heat loving) bacteria. Until recently ...only available Cas9 proteins were isolated from mesophilic (medium heat loving) bacteria, such as Streptococcus...by the Doudna lab has opened up the field of thermophilic bacteria to CRISPR/Cas9 genome editing. How...Harrington et al discovered a Cas9 protein from the thermophilic bacteria Geobacillus stearothermophilus (GeoCas9...could also be used with in vitro molecular bio applications that require high temperature reaction conditions...

...other fun activities and even invite members of the public to attend. We thought we’d take some time to introduce... University of Cambridge and within the general public. The society organises talks by prominent scientists...sessions to help undergraduate students with their applications. The society currently has 1,283 members, who...students, postdocs, faculty members, and the general public. Chemical Society in University College Dublin,...range of talks for both students and the general public, career events, and networking meetups. The society...