...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 ...

...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...

...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...

...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...

...targets to measure! Let’s take a look at some of the hottest fluorescent biosensors new to Addgene this year...year. Feel the love with MTRIA sensors One of the latest additions to Addgene’s catalog is a new class of...smell the flowers - or rather, the lactate! The latest biosensor from Gary Yellen’s lab is LiLac, a fluorescence...fluorescent proteins with different thermal quenching rates (mNeonGreen and tdTomato), they produce a temperature...over previous temperature indicators, it also demonstrates that taking advantage of fluorescence quenching...

... used to identify a gene mutation is the genetic test used for sickle cell anemia (2). Southern blotting...of restriction enzymes that cut DNA at specific sites (5). Imagine that you have a genome with unknown...cloning and analytical digests. A researcher can test the orientation of an insert in only a few hours...available although they are reported to have high error rates (10). It is projected that nanopore-based sequencing... C., and Yuet Wai Kan. "A sensitive new prenatal test for sickle-cell anemia." New England Journal of ...

...recombination events, and the synthesis of alternative loxP sites to specify distinct patterns of recombination. In...Molecular and Cellular Biology, harnessed these attributes of Cre-lox to create the Brainbow mouse neural...defined using 2 loxP variants (only identical loxP sites can mediate recombination.) The deletions are mutually...be detected via immunostaining. This feature facilitates screening of Cre-negative Brainbow mice to determine...doi: 10.1038/nrn2391. Epub 2008 Apr 30. PubMed. Intestinal crypt homeostasis results from neutral competition...

...with a red-shifted fluor. Here, NLuc internally excites the fluor through a process known as bioluminescent...exogenous excitation and with OgNLuc being the brightest since it had an emission near 600 nm – a clear...imaging. In all of these fusions coelenterazine intiates the fluorescent signal and these FP-RLuc8 fusions...see the movies demonstrating Ca2+ dynamics in cardiomyocytes. Enhanced Nano-Lantern constructs are available...bright cyan-excitable orange fluorescent protein facilitates dual-emission microscopy and enhances bioluminescence...

...there is variable transduction of oligodendrocytes, microglia, astrocytes, neurons, and inhibitory neurons...Serotype Oligodendrocytes AAV8 > AAV2 & AAV6 Microglia AAV9 > AAV2 & AAV6 Astrocytes AAV8 > AAV2...previous literature using AAVs for DREADD delivery indicates ~7-21 days are required for the onset of gene ...

...curated their Ebola-related content. -And the latest updates on the ongoing outbreak from the CDC and WHO...Dallas was the first to be infected in the United States while caring for a patient, despite wearing protective... data on five strains of Ebola representing the latest outbreak and earlier ones, along with links to ...

...References: Skeletal muscle PGC-1α1 modulates kynurenine metabolism and mediates resilience to stress-induced...up in mice as poorer performance on forced swim tests and less interest in sweets. Further examination...be important to identify the key proteins and metabolites that are dynamically secreted by muscle during...

...double-strand breaks (DSBs). DSB repair by HDR facilitates in-frame insertion of tag-encoding sequences ...assembling csRNPs in a 96-well plate format and tested parallel tag knock-in for 96 different genes in... negative control for the tagging experiment and test them in parallel so you have a good measure of ICC...clonal lines by single-cell deposition in 96-well plates: Characterize clonal lines by ICC and PCR genotyping...any cells positive for tag insertion, you should test a new crRNA and/or try out an alternative tag (3xFLAG...

...discovery. With these screens, the investigator generates a cell population where all genes are knocked ...groups and, as a consequence, which genes are good candidates for being involved in the studied phenotype. ...These targeted screens can allow a researcher to test which of the differentially expressed genes found... is added), and decide which genes you’d like to test for phenotypic effects. Typically the phenotype ...interest (that were not found by Green Listed), and test if any of the alternative names works better. To...

...costume, and did their best to claim the costume contest trophy. We’ve had teams dressed up as plasmids,...table. This year, in addition to the Halloween contest, we’ve also carved pumpkins (yes, Blugene made ...temperature and humidity favors fungal growth. The ants bites down and locks their jaws into a leaf, and dies ...Addgene See some of our previous Halloween costume contests ...

... (see schematic below), enabling a researcher to test multiple tags for one protein quickly and easily...easily. These tagged plasmids also include cleavage sites should one need to remove the tag post-purification...post-translational modifications added to proteins in other eukaryotes better than E. coli. Plasmids designed for these... systems may include a signal peptide (SP) that routes the protein through the secretory pathway to improve...