...regions flanking the target site).  The best applications for dsDNA inserts include: large insertions ...targeted, it’s best to determine the direction of replication through your region of interest and design an...it’s easier to not determine the direction of replication and instead design oligos that target both strands...correct DNA mismatches that occur during DNA replication. There are two ways to avoid activating MMR: ...Uses in Recombineering Substrate Best Suited Application Recombination Frequency Special Considerations...from a plasmid with a heat sensitive origin of replication.  Once the recombineering system is no longer...defective prophage is to use mini-λ, a defective non-replicating, circular piece of phage DNA, that when introduced...

...tropism, replication kinetics, and cell damage profiling of SARS-CoV-2 and SARS-CoV with implications for ... Another example of reagents that have broad applications but have proven useful to the study of SARS-...vectors VSV-G and psPAX2, which were used in a publication by Xiuyuan Ou, et al. that characterized the ... requested these plasmids from Addgene. This publication then resulted in the development of a new plasmid...scientific community. Deposits of new reagents, policies from journals and granting agencies requiring ...

...objects that exist on the internet so that when you click on the link it will take you directly to the original...item. Given a specific Addgene ID # listed in a publication, you can quickly pull up the associated plasmid...plasmid page also helps researchers find other publications that have used the same plasmid through the ...section. Having IDs linked to these metadata and publications help to enable reproducibility of experiments...and N2T.net. To help authors comply with journal policies, Addgene now provides an RRID and a resolvable... in the material and methods section of your publication. You should carefully read and follow the journal...

...2020.  “CRISPR Gene Therapy: Applications, Limitations, and Implications for the Future.” Front. Oncol... for full details.    CRISPR advances in monoallelic diseases In 2013, a team led by Jinsong Li from...the technology in different labs for different applications.” Li’s team noted that CRISPR/Cas9 has been ...with size limitations. In addition, non-viral applications are also possible through the use of nanoparticles....  “CRISPR Genome Editing Technology and its Application in Genetic Diseases: A Review.” Current pharmaceutical...

...together a grant application? Are you familiar with the publishing or patent application processes? How ... pages in a Curricula Vitae to showcase your publications, academic performances, and research experiences...information. Think "skills," not just "research and publications” What are the skills you developed through your... science? Or perhaps you have presented in some public lectures? These transferable skills could be meaningful...make their job postings and salary expectations public, so start by looking at these university postings...

...your application a whole week before the deadline. The day of submission, review your application with... their dream.  To help students focus on the application process, and simplify access to academic resources...Scrutinize every opportunity prior to starting an application to assess your eligibility, interest, and career...February represent peak season for internship applications. Thus these noble individuals are inundated ... with a stellar letter of support.        5. Application submission  Set a calendar reminder for every...

... our knowledge. Disease Applications Q13: Does AAV have potential applications in diseases which may be... to use split-inteins to mediate trans-protein splicing. Note, however, that this will require co-delivery...reliable and reproducible. However, we do not use amplicons in ITR as we have found this to strongly bias ...1x1014 vg/ml.. As evident in our repository, this application is an outlier for PHP.eB (1x1011 - 1x1012 vg/...CRISPR tools is an exciting area for future AAV application to epigenetic editing. This area is discussed...

...protein for your application. Selecting the best fluorescent protein for a specific application can seem like...under conditions that best mimic the intended application. The three key properties for live cell imaging...experimental set up. The practical brightness takes all application specific parameters into account, including the... picture of what can be expected in a 'real' application than the theoretical brightness. Consequently...brightness in a system closely mimicking the future application. In figure 2 we provide an example of the comparison...

...transcription, or DNA repair, or replication, or any number of CRISPR applications and thought “OK, but how does...with Addgene, but you’ll find many great tools by clicking on the links below and visiting their associated...containing lists of plasmids from the indicated publications. You can request all of these plasmids directly..., Hoefler R, Schmalhorst PS, Jude JG, Zuber J, Gerlich DW Cell. 2017 Aug 24;170(5):956-972.e23 Screen ..., Hoefler R, Schmalhorst PS, Jude JG, Zuber J, Gerlich DW Cell. 2017 Aug 24;170(5):956-972.e23 Rapid and...

...tool that can be used for a variety of research applications. Within the immune system, antibodies are produced...antibodies to bind proteins is useful for research applications as well because they allow scientists to target...150 kDa, a nanobody is 12-15 kDa.   Research applications for antibodies You can use antibodies for experiments...Let’s dive in and take a look at some of these applications. Western blot Western blots are used to detect...immunocytochemistry). This could be cells or tissue slices mounted to a slide, or whole organs. Because the... by binding to cell surface receptors. These applications are typically used for investigating signaling...are out there, and how many different research applications they can be used for. Addgene’s Antibody Plasmid...

...ampicillin resistance cassette and an origin of replication, which are not schematized. Sizes of fragments...2024, used under the Creative Commons CC BY-NC-ND license.   Once you have the components you want, simply...validated and were chosen because of their broad applicability in different model systems. What’s the error...five months. When they tested those clones (in duplicate), 93% had the correct banding pattern in the restriction...correct. Because they created the assemblies in duplicate, they were able to confirm that at least one plasmid...design. Note that you can browse all components by clicking the blue text in the upper right-hand corner.  ...

...chemical replication of RNA in the absence of evolved enzymatic machinery (i.e. RNA replication with no...technology park, it enjoys preferential governmental policies for a high-tech development zone so many science...inheritance that might be less error prone than DNA replication. He too mentioned how “surprising” small RNAs...created a circRNA website so scientists can view public circRNA datasets and download the custom python...Steitz quipped that RNA in the ribosome is in a complicated conformation “not like DNA which has a boring...

...are, you’ll even have DNA left over for other applications, like PCR, cloning, transfection, or long-term...wash step or use a different strain. For most applications, Addgene uses and recommends DH5𝝰! This strain...your E. coli to ensure it is suitable for your application. We have previously reviewed many common lab ...partially specified by a plasmid’s origin of replication, but can also be affected by the plasmid size...the material used for inoculation. To further complicate matters, plasmid copy numbers are often ambiguous...volume limits, the copy number of some origins of replication (e.g. p15A, ColE1) can be increased by adding...

...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...

... 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...

...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...