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... development of several fluorescent autophagy reporters now allows for the measurement of autophagic flux...makes Keima useful for monitoring mitophagy. It’s important to remember that because Keima’s fluorescent relies... al. "Rosella: a fluorescent pH-biosensor for reporting vacuolar turnover of cytosol and organelles in...using a single fluorescent biosensor." Scientific reports 5 (2015): 12397. PubMed PMID: 26215030. PubMed ...

...reaction Run PCR to amplify your insert DNA. It is important to use a high fidelity polymerase to minimize .... The fidelity of the polymerase becomes more important the longer the expected PCR product is. You should...temperature based on the melting temperature (Tm) of the portion of the primer that hybridizes to the sequence to...used. No matter which polymerase you use, it is important that you sequence the final product. Additional...

... cells and activate expression of a luciferase reporter. CPP was not required for protein entry into cells...primary cell types using Cre and various loxP reporters. In both mouse and human embryonic stem cells,...indicating a 70% frequency of biallelic mutations. Importantly, hESCs retained their stemness, indicating that...Plasmids @Addgene: iTOP-tested proteins and reporters Previous Protein Delivery Techniques: Introduction...

...finding mentors and, perhaps, asked someone to support your career development as your mentor. How do ...mentoring relationship really productive, the most important step you can take is to set goals to work on together...to help a Mentee reach goals. One of the most important contributions made by a Mentor is to help the ...your creative thinking Enhance leadership skills Support inclusion, diversity, and open communication Feel...

...will include, but is not limited to temporary housing, transportation, living expenses, and necessary reagents...visits an institution to perform research there temporarily. The length of stay could be from a month to ...forget to have fun! Visiting is an amazing career opportunity, but also a really good time! If it sounds right...

...cerevisiae to human: The important gene co‑expression modules. Biomedical Reports 7.2 (2017): 153-158. DOI...attention for its powerful promoter of a gene important for methanol usage. This gene promoter, AOX1, ...., Comparative genomics of biotechnologically important yeasts. Proc. Natl. Acad. Sci., 113, 9882-9887...

...SARS-CoV-2 were developed for broad use and are now important for accelerating research into this virus. One...during the pandemic and in the future, depends on support from the scientific community. Deposits of new ... be deposited into repositories, and financial support to maintain repositories’ capacity for storing,...all ways in which the scientific community can support repositories. Repositories are essential for the...

..., the MIT Technology Review dropped a bombshell report. A scientist working in China was using CRISPR/...While the Nuffield Council has recently published a report on the social and ethical issues of genome editing...question of “how do we move forward” is the June 2012 report on the ethics of “Novel techniques for the prevention...diseases. The key success of the Nuffield Council’s report is that when it became clear that this assistive...

...back in with the Addgene blog for future award opportunities. Xin Zhou: Fluorescent Light-inducible Proteins...particularly notes their ability to be precisely spatio-temporally controlled as a key advantage over other inducible...Oligosaccharides (HMOs). HMOs are thought to play an important role in nuturing the development of beneficial...consisting of sfGFP fused to a His-tagged ABC transporter known to bind to the HMO of interest. Fatima ...

...ApoB-NanoLuc reporter by inserting the NanoLuc luciferase downstream of the apoB gene. Using this reporter, the...Cell, 2019. PubMed PMID: 30639098. LipoGlo: A reporter for measuring ApoB bound lipoproteins Article ...cells. These changes were larger than previously reported for other organisms, and the Gottschalk group ...

... 91bp on the PAM-proximal side, consistently supported higher frequencies (up to 60%!) of gene editing... and the targeting guide RNA into cells by electroporation. Results from our lab and others (Lin et al...Kim et al 2014) have shown that this technique supports extremely high frequencies of genome cutting and...nucleofection as (in our hands) this technique supports gene disruption or gene replacement frequencies...

...Nature has evolved ways to prevent degradation of important cellular RNA molecules with post translational...your gRNA. Pro tip: modifications are typically incorporated at the termini of the crRNA molecule (usually...Frontiers in Genome Editing). This is particularly important in experiments in which viability is being compromised...Ramel, MC., et al. Photoswitchable gRNAs for Spatiotemporally Controlled CRSIPR-Cas-based Genomic Regulation...

...about the process that was used to create this important collection.  Creating the Plasmids To begin constructing...antibody (R-mAb) expression plasmids, allowing for important reagents that could have been lost to continue...sequences available from each plasmid webpage. Importantly, the methods developed by the Trimmer Lab for...why choosing the right secondary is of primary importance. PLoS One. 2012;7(6):e38313. doi: 10.1371/journal.pone...

...permanently integrate into the host genome or temporarily express whatever it encodes. This ability to ...straightforward across applications, while electroporation of a human, for example, isn’t a reasonable...allows you to infect cells and deliver cargo, it’s important to understand their differences to choose the ...PMC413431. 2. Bukrinsky MI, et al. “Active nuclear import of human immunodeficiency virus type 1 preintegration...

...severely cold conditions. To understand their unique importance, let’s begin with a few basics.  Mesophiles vs...understanding and appreciating their biology. The importance of psychrophilic enzymes From an industrial perspective...decided to create a hybrid system where we could incorporate cold-tolerance inducing genes within a regular...scientist from Addgene. The talk highlighted the importance of reagent repositories and the myriad facilities...

... phage promoter pR, preventing expression of a reporter gene. When light is present, YFI is inactive, ...preventing repressor cI synthesis and allowing reporter gene transcription to take place. Addgene also...promoter is right for me? Here’s a list of some important variables to consider when choosing an inducible...’re expressing a gene that may be toxic, it’s important that your inducible promoter not be too leaky....

...most important resource that Addgene provides the scientific community? Katz: The most important resource...beyond. In your perspective, what do you think are important considerations for the future of open science?...immensely. Paratore: I have two children active in sports, so I enjoy watching their games.  If I’m not at...

...binding ability of its nanobody partner, so it’s important to insert AsLOV2 at a spot in the nanobody that...light to control signaling provides greater spatiotemporal resolution than pharmacological or genetic ...activated Ras/Erk signaling by using a fluorescent reporter for Erk kinase translocation. These results demonstrate...control protein activity Read about RANbodies, reporter nanobody fusions Learn more about using the secondary...

...nuclease for your experiments, there are three important factors to consider: site accessibility, specificity...potential for off-target cleavage should be an important consideration during any experiment. Sensitivity...stringent PAM specificity. This can be especially important for base editing techniques, where Cas9 must be...many options! Comic by Maya Kostman. Another important consideration when choosing a Cas9 is balancing...

...scientific conferences. Conferences provide the opportunity to meet people who are interested in the same...the conference. Large conferences have many opportunities but can be overwhelming as far as meeting new...the conference The conference may start in the airport. If you see someone traveling with a poster roll...friendly so stick with those. Don’t miss social opportunities, but don’t overdo it (some of my best studies...