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...cells, and has potential use for a wide range of applications in cellular biology. Van Haren J et al. Nature...versatile in vivo fluorescent reporter for rapid quantification of base-editing and Cas9 cleavage activity ...plasmid that allows for rapid fluorescence-based quantification of base editors that utilize dCas9 fused to... The reporter constitutively expresses eGFP to indicate reporter abundance, and mCherry fluorescence is...frame-shift mutation within the mCherry reporter. Quantification of editing efficiencies simply involves tracking...

...features such as antibiotic resistance markers and replication origins, but there is extensive heterogeneity...microbial strains. It proved to be impractical to catalog every variant of a feature. Instead, we identified...occasional mismatches or indels. Empirical tests indicated that a reasonable rule is to require at least ...that have different properties. To prevent misidentification, we augment the database with new fluorescent...protein variants. The future of DNA feature identification What’s next for common features? We plan to...

... (AmpR) and origins of replication (ori and f1 ori) which allow it to replicate.   How can I prevent...the bacterial selection marker and origin of replication. During bacterial culture, only the plasmid with...the antibiotic resistance gene and origin will replicate, and the transgene will be lost. Over time, bacteria...the recombined plasmid is still visible. This indicates that the DNA prep contains a mixture of the full...

...containing only the sequences required for viral replication and transcription as well as reporter genes. ... membranes. Read the full post for potential applications of R-bodies and to find a video of them in action...Medical School describes how AAVs function and indicates important things to consider if you’re getting...these libraries from the post and the associated publication. Optimizing Donor DNA for Enhanced CRISPR Genome... to your DNA sequence of interest. Potential applications of the system include activation and repression...

... and testing so that researchers can use our educational resources and experience to their advantage should...preparations have worked well in my lab, including in applications where we have delivered them in vivo to the ... help! We’ve been expanding our viral vector educational resources as well, with both posts on the blog...distributing new types of materials. At the time of publication we’ve already fulfilled more than 140 orders ... service as we continue to develop it. More communication and sharing will only help research advance....

...template is supplied. Find Plasmids from the NgAgo Publication at Addgene Cas9 and NgAgo head to head NgAgo ...differences in cleavage efficiency. If this finding is replicated in larger studies, it will represent a key advantage...site specified by the gDNA. Depending on your application, this could be favorable or unfavorable. In a...PubMed PMID: 27136078. Find plasmids from this publication at Addgene. 2. Swarts, Daan C., et al.  “DNA-... PMCID: PMC4697943. Find plasmids from this publication at Addgene. 3. Swarts, Daan C., et al. “Argonaute...

...Neurobiology has been working on genetically encoded indicators of calcium and other small molecules since the...lymphocytes, too, making them a versatile tool for application in both neuroscience and immunology. Don’t be...imaging for beginners Griesbeck says his calcium indicators can be used in mouse, Drosophila, C. elegans,...give it a shot. “Beginners sometimes think it’s complicated, but actually it is very good,” he said. “It ...his team is now applying it to improve upon an indicator they developed and published in 2010 for visualizing...

...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... 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...

...processing, these small DNA strands can form concatemers in later steps. This new NEB kit includes a step...fragments the DNA and prevents it from forming concatemers later. The "FS" stands for Fragmentation System...repair Adapter Ligation Clean up/Size selection Amplification Clean up  I found this kit easy to use because...DNA we started the process with, we got great amplification of our sample libraries. NEB also gave us a ...mostly used to make new libraries as opposed to replicating them like we do at Addgene), going to the workshop...

...-guide RNAs – but it also allows you to skip purification steps and end-repair steps with well-designed...that the region of homology doesn’t have to be located at the absolute end of a fragment. This means that...fragment sequences and the polymerase used in the amplification. This tool allows instant updates as fragments...transforma­­­tion or as a PCR template. Some of the applications of the NEBuilder HiFi DNA Assembly Mix include...the limitations associated with otherwise more complicated assembly approaches, and opens the door for new...

...researcher and science communicator.  Update (7/17/2018): Check out Fatima's recent publication on her HMO work...protein-protein interactions. Beyond these common applications, researchers are finding new ways to apply fluorescent...utility of these FLiPs in a variety of research applications and particularly notes their ability to be precisely...endeavors, Fatima hopes to practice her science communication skills throughout her training and wants to ...

...Marketer Software Developer Learn Science Communication Skill from a Student Group Like Science in the...regular travel be required or will you stay in one location? How long is the commute? Will the job require...online job websites to see what requirements or qualifications are needed for the role, as this will give ...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...

...ll be talking about the ins and outs of this classification system, including its history, biology, and...repeatable phenomenon, Gram began using it as a classification system. Eventually, any bacteria that retained...permeable to different classes of antibiotics. The classification system is also useful in a laboratory setting...bacteria type. The gram positive and negative classification system is incredibly helpful in visualizing...stepping into a lab.   Anna Kamens is a science communication summer intern at Addgene and a rising third-...

...transcription, or DNA repair, or replication, or any number of CRISPR applications and thought “OK, but how does...pages containing lists of plasmids from the indicated publications. You can request all of these plasmids ...below pull up lists of plasmids containing the indicated gene, and you can subscribe to these pages to ...

...for pre-clinical and clinical applications of dCasRx. The identification of Type VI Cas13d enzymes is ...RspCas13d. RspWYL1 also increased EsCas13d activity, indicating that WYL domain-containing proteins may be broader...even when guides were tiled across a target RNA, indicating that this enzyme does not cleave at a predictable...target essentially any RNA in a cell. Since catalytically dead dCasRx maintains its RNA-binding properties...

...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...

...cohesive team environment. Neglecting communication skillsClear communication is the backbone of effective management...management. Miscommunication often arises when managers fail to set expectations or provide timely feedback...firsthand when it took me some time to adapt to the communication style of a new, junior team member who had values...and leveraging tools like the Myers-Briggs Type Indicator or DISC assessments can help you navigate team...

... our knowledge. Disease Applications Q13: Does AAV have potential applications in diseases which may be...Did you catch our April AAV webinar with Tim Miles, PhD, Director of the CLOVER Center at CalTech? If...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...

...of the biosensors used in research are very sophisticated, but at their core they can all be boiled down...ready-to-use. While biosensors span the gamut of applications within chemistry, biology, and physics, in this...sensors can be targeted to different cellular locations or tissues depending on the biology you are studying...different factors, in different or the same cellular location, all at once! Small molecules What type of small...Neurotransmission and Neuromodulation: Engineering and Applications. Frontiers in Cellular Neuroscience. 2019 Oct...

...thus allowing repair to take place before DNA replication. Indeed, they observed that 41/42 embryos were... embryos carried indels at the mutation site, indicating that repair proceeded through NHEJ rather than...attributable to CRISPR editing. Conclusions & implications One striking feature of this paper is the failure... need to be much, much lower before clinical applications could be considered. It is important to consider...experiments are needed before we can consider clinical applications for germline editing. Although this paper represents...