...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...

...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...

... 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 ...

...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 ...

... 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 ...

...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...

...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 ...

...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...

...genes, which code for polymerases, nucleases, and helicases, were also disrupted to elucidate their role in...2013 (7, 8). Nine months later, more than 1500 publications describe work to improve the tool’s specificity...various species and describe an assortment of applications. Despite the ubiquity of CRISPR/Cas toolkit,... multiple plasmids used for various platform applications. For more information on CRISPR/Cas, check out...for your research. Do you have a cool idea or application that involves CRISPR/Cas technology? Let us know...

... guest bloggers Aline and Benjamin Glick from SnapGene!  Ben Glick is President and Chief Scientist at...contribued by guest bloggers Aline and Benjamin Glick from SnapGene. SnapGene was created to meet a need...features such as antibiotic resistance markers and replication origins, but there is extensive heterogeneity...because he needed it for his own lab.     Aline Glick is VP of Product Management at SnapGene. She is ...

...permission from the Genetics Society of America. Applications of this system Dickinson et al. have developed... in cell culture. In addition to CRISPR/Cas9 applications, SEC selection could replace other, more restrictive... of print] PubMed. Find plasmids from this publication at Addgene. Other strategies for CRISPR/Cas9...2013 Sep 1. PubMed. Find plasmids from this publication at Addgene. Scalable and versatile genome editing...2014 Sep 23. PubMed. Find plasmids from this publication at Addgene. Additional Resources on the Addgene...

...critically and carry out in-depth research is applicable to a wide range of industries and is a highly... How long is the commute? Will the job require public speaking? Will you be leading a group or working...sectors such as forensics currently have too many applicants compared to the number of available roles). Make...your research, it’s time to start sending out applications. Check out our other career posts for more specifics...careers and to find advice on crafting the perfect application. Additional Resources on the Addgene Blog Learn...

...learned about from textbooks just made everything click in my head and left me thinking: I wish I had this... expensive tiny amounts of reagents are, how complicated (and again, expensive) the different machines...proteins without the need to handle a living, replicating organism. Furthermore, cell free systems can ...PMCID: PMC6070313. Find the plasmids from this publication at Addgene! Additional Resources on the Addgene...pages Find plasmids from the BioBits BrightTM publication here ...

... Overlap extension PCR is used to splice together two PCR amplicons based on sequence homology, and the...mutations, large deletions/insertions require complicated, often costly in vitro assembly methods. A new...appropriate vector. Gibson Assembly, GeneArt Cloning, and SLIC can all be used to assemble multiple fragments, ...Read about Other Plasmid Cloning Techniques like SLIC and Golden Gate Cloning Learn How to Verify Your...

...resulting in a fluorescent signal. Since their first publication in 2012, the Nagai laboratory has assembled a...multicolored nano-lanterns for use in various applications, including optogenetics, biosensors, and fusion...sensitivity and faster imaging compared to previous publications. They also developed calcium, cAMP, and ATP ...furimazine. In addition to the previously described applications, the enhanced signal of eNL allows their use...characterizing Nano-lanterns and eNL for many different applications. For long-term imaging, especially where photobleaching...

...7). Ability to distinguish between mono-allelic and bi-allelic mutations: Perhaps the most noteworthy ...derived samples with mono-allelic mutations versus those with the bi-allelic mutations typically desired...a gene of interest is one of the most common applications of precision genome editing, the remainder of...

...function relationships or, in more practical applications, allow a protein to be more effectively used...the creation of many new tools and potential applications. But this focus on development, along with the...’t give it a try. Optimization Why is GCE so complicated? Well, while the techniques are straightforward...has “10x the impact of PCR. But it’s 100x more complicated.” And Ryan Mehl would know. He’s the director...ncAA protocol, to optimize and share, they’re soliciting feedback from scientists on which protocols would...

...convinced that most microscopy techniques can be replicated with commercially available consumer-grade components... tools has allowed the community to regularly replicate designs worldwide. Distributing hardware via globally...the construction plans. Appropriate open-source licenses for hardware (e.g. CERN OHL), software (e.g. GPL...questions to be clarified, especially with regard to licensing models. Direct sharing of work from universities...universities and research centers provides a potential conflict between traditional IP-based models and the use...

...As evidenced by all the CRISPR publications, press, and plasmids out there, it’s obvious that CRISPR ...they have a higher propensity to reform double helices and decrease Cas9 cutting efficiency. However, ...PubMed PMID: 26628643. Find plasmids from this publication at Addgene. 2. Nishimasu, Hiroshi, et al. "Crystal...PubMed PMID: 26735016. Find plasmids from this publication at Addgene. 5. Chen, JS, et al. "Enhanced proofreading...PubMed PMID: 28931002. Find plasmids from this publication at Addgene. Resources on the Addgene Blog Read...