...mechanism of their voltage sensing component, which separates them into two distinct categories detailed below...phosphatases from sea squirts. This design structurally separates the two components of the indicator, which can...was dipicrylamine, a lipophilic compound that distributes itself within the plasma membrane as well. Another...particularly physiologically relevant in neurons, myocytes, and endocrine cells. It follows that the specific...component (i.e., when the dye is lipophilic and integrates into the membrane), the voltage sensing component...

...quality control, but does not perform functional tests. The newest genome engineer sharing the stage with...be less desirable if one has to target multiple sites in a single experiment. Also, the NGG PAM sequence...nearly 400 times. Researchers have been excitedly testing NgAgo's genome editing applications. So, how does...conducted a survey and asked researchers if they had tested and compared NgAgo head-to-head with CRISPR-Cas9...could not achieve indels with the NgAgo system indicates that it is very tricky to use and fussy to optimize...

... plasmid with multiple, individual expression cassettes, having the genes expressed from the same cassette...described below. IRES Elements Translation in eukaryotes usually begins at the 5’ cap so that only a single...originally discovered in poliovirus RNA, where it promotes translation of the viral genome in eukaryotic ... start with a plasmid that has multiple cloning sites flanking the multicistronic element(s), or you could...efficiency of a 2A peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice. Kim...

...biological questions. The two most common uses are: 1) to test the expression level in a specific system (by measuring...occurs when energy transfer from one FP (e.g. CFP) excites the fluorescence of another FP (e.g. YFP). FRET...create the chromophore. This can be from a few minutes after it is translated to a few hours. For example...However, for most FPs it is a few seconds to a few minutes. Photostability can be affected by experimental...greenfluorescentblog.wordpress.com.   More Helpful Websites & Resources: FP guide at Addgene Interactive ...

...cell types. Variance can be caused by cell cycle rates; rapidly diving cells will have higher HR frequencies...viability of your cells. Enrich S-phase cells HR operates predominantly in S and G2 phase of the cell cycle...carefully (Lin et al., eLife)! Use a Cas that facilitates HR What if promoting HR was as simple as swapping...It can be! Cas12a is a Cas family member which generates sticky ends upon cutting. Cas12a may promote HDR..., Rouet, R., Vejnar, C. E., et al. CRISPR-Cpf1 mediates efficient homology-directed repair and termperature-controlled...

...insert-specific sequence. This is a “yes or no” kind of test, with a positive clone amplifying a product and ...primers. These primers are designed to anneal to sites that flank the insert site. A positive clone will...depends on your preferences. Either way, make sure to test your colony PCR primers before using them to screen...Pros Cons Insert-Specific Simple "yes or no" test results Doesn't tell you the orientation of ...of sterile water.  Pick 3-10 colonies in total to test, depending on the number of background colonies ...

...visual appeal, instant sharing across multiple other sites), I created a new platform for sharing video abstracts...communicating research. Short videos of 3 to 5 minutes provide more than enough time for researchers to...website that helps researchers go beyond citation rates to measure the impact of their contribution. It ...Vimeo, and other platforms (such as university websites). WeShareScience is a social platform that provides...about her research on plummeting embryo survival rates. Video on We Share Science: http://wesharescience.com...

...initiative and start organizing meetings and take minutes. Get to know your collaborators from other institutions...LinkedIn or ResearchGate allow you to send personal updates as messages and keep you up-to-date on your collaborators...collaborators for example on a holiday or for professional updates. Whenever I see interesting events, articles and...make sure to share them with a short note - this creates a thoughtful way of interacting and is much more..., checking in regularly, and maintaining useful notes that remind me about the current state of a partnership...

...to get involved in product testing in the lab, to travel to customers’ sites, and to run experiments with...for a high grant success rate. At this point, candidates are highly experienced but often have to start...requiring in-depth analysis, data review, in-house testing etc. Technical support roles are primarily office.... Problem solving and analytical skills are prerequisites but these too can be enhanced on the job. When...

...back to) is the one-eyed pinhead mutant, which regulates a developmental pathway implicated in congenital...Zebrafish Model Organism Database ZFIN actively curates information on mutants and other resources. Targeted...oligonucleotides called morpholinos. When injected into zygotes, morpholinos complementary to a target mRNA inhibit...help elucidate the gene networks that transform zygotes into adults and explain how signaling pathways ...study human illnesses such as blood disorders, diabetes, cancer, and Alzheimer’s disease. Naturally, the...

...quickly screen and investigate transcription factor candidates and the regulation of genes in the biosynthetic...save you a lot of time. Don't re-invent the wheel. Test different leaf ages Usually younger leaves work ...interest. Find all plasmids from the Lee-Parsons lab. Test different leaf infiltration methods Co-cultivation...transcripts in the plant but not from Agrobacterium. Test different Agrobacterium strains We used the disarmed... This allows you to quickly assemble vectors and test different ideas. We are a huge fan of the Golden...

...nil base editing efficiency at some genomic sites. Some sites may be inaccessible to base editors probably...cytosine deaminase with Cas9 nickase (nCas9). nCas9 creates a nick in the DNA by cutting only a single strand...indel formation. After binding with DNA, the CBE deaminates a target cytosine (C) into a uracil (U) base....Gaudelli et al., 2017). Our vectors have two Bsa1 sites to clone the single guide RNA (protospacer+scaffold...

...country.   Figure 1: Number of countries Addgene distributes to per year, 2005-2023. We identified and offered... we contracted with so we can always offer the fastest shipping. However, there were issues that persisted... country are represented in blue; darker blue indicates more views from that country.   While written ... country are represented in blue; darker blue indicates more views from that country. Stats and figures...resources shared via social media, on collaborators’ websites, or as a recommendation in a Reddit thread crowdsourcing...

...chemiluminescent substrates are available. Labs with limited equipment often opt for the chromogenic substrates since...antibody buffer components can interfere with conjugates. Make sure to do your research before conjugating...sandwich ELISA. In this method, the target antigen competes with a reference antigen for binding to a primary...is non-specific binding or false positives.  If testing a recombinant protein, especially one with tags...

...factor) promotes editing (Yan, J et al., 2024). They developed a new prime editor that incorporates La’s ...AAV Vector System Neurodegeneration Collection updates   Multiplexed perturbation and decoding in pooled...Figure 6: Illustration of split fluorophore assay to test reconstitution via mRNA trans-splicing. A cis-splicing... New and notable Neurodegeneration Collection updates Last but not least, check out the new and noteworthy...

...typically contains iterons that serve as recognition sites for replication initiation proteins. These iterons...The plasmid does not have many restriction enzyme sites. Hosts often have a restriction barrier as a defense... al. 1977). So if a plasmid has few restriction sites, it may be able to sneak past this checkpoint.  ...incorporate interchangeable antibiotic resistance cassettes (Prior et al. 2010), inducible promoters with ...inducibility (Toh et al. 2023), and additional cloning sites (Keen 1988).  Find broad host range recombinant ...

...PE6b was very efficient for short, unstructured templates and significantly reduced undesired edits (indels...complex edits, such as long or highly structured templates. Cas9 variants PE6e–g often had even higher editing...transcriptional histories, accurate to within 40 minutes. Figure 4: Time-resolved protein ticker ...-PSD-95 [K28/43R] recently passed their initial tests with thick slice (70 μm) immunohistochemistry (IHC...fact, Anti-PSD-95 [K28/43R] has been one of the hottest antibodies in Addgene’s collection since we began...

...scientific mentor is typically a professor who operates a research group. Within that research group, ...mean everything! The bare minimum should be taking notes on general protocol steps and other tips and tricks... in the next few days or weeks), look over your notes and protocol(s). Is there enough information there...throughout training, while others prefer to review their notes, process what they’ve just learned, and ask questions...for a copy of the protocol if you prefer to take notes on a printed protocol, or maybe you would like to...

...and dates of your actions. These records can take the form of written comments, added files, notes etc...Cancer Biology have shown similarly disappointing rates of reproducibility. These alarming results are due...the morning, to the daily news briefing, to quick notes, to finances, to flights & ordering food - everything...everyday work to capture ideas, write down certain notes or brainstorm, but we have to admit that paper-based...

...binding of an aptamer to its target small molecule activates the fluorescence of the small molecule. Fluorescent...occurring regions in the 5’ UTR of mRNA that bind metabolites, leading to conformational changes that control... metabolite binding to the riboswitch instead activates Spinach fluorescence. In the search for something... to identify aptamers capable of inducing the brightest fluorescence (Filonov et al., 2014). Broccoli ...2017). Corn naturally dimerizes, and this dimer activates the fluorescence of 3,5-difluoro-4-hydroxybenzylidene-imidazolinone...