... in bacteria CRISPR-Casɸ mGold, a new yellow fluorescent protein New CRISPR plasmids New items from the...32675376/ mGold, the most photostable yellow fluorescent protein to date By Justin Ng High-throughput ...High-throughput screening methods, such as fluorescence-activated cell sorting (FACS) helps researchers sift through...million cell types expressing variants of Yellow Fluorescent Protein (YFP). The team identified a new YFP ...Resources page.  CDDR (CRISPR–Cas9-based Dual-fluorescent DSB Repair) is a double-strand break repair assay...the introduction & resolution of two DSBs in a fluorescent reporter. Find the plasmids here Yongsub Kim...

...study a wide variety of in vivo conditional gene rescue experiments, as well as for improvement of optogenetic...additional uses for FLEx switches: Conditional rescue of a gene knockout: To test if the phenotype you...by coding sequence of the gene you would like to rescue with in the antisense orientation (let’s call the... to test whether or not expressing GeneR alone rescues the phenotype being studied. Introduction of ...functional mutation in vivo: Similar to a conditional rescue experiment with a wildtype copy of a gene, you ...presence of Cre, GFP expression is lost, allowing for rescued expression of GeneR.   Sometimes also referred...

...plasmid, how will it be sent to me? Description: This describes how the plasmid or other product (kit...useful to me? Plasmid purpose: This is a brief description of what the plasmid was designed and used for...vector’s plasmid number here. Gene/insert: This describes what has been inserted into the backbone, and ... sequencing. Growth in Bacteria: This section describes how to grow the bacterial strain in which the ...associated, such as when a plasmid contains certain fluorescent proteins. Finally, for many plasmids you can ...

...Western blots. ELISAs. Immunofluorescence. What do all of these techniques have in common? They all typically... distance between the target antigen and the fluorescent label, which can reduce the potential image resolution... resolution of super-resolution fluorescence microscopy. Their size also limits their ability to penetrate...penetration and decreases the distance between a fluorescent tag and the target antigen to about 2 nm, which...review. If you just want to use the nanobodies described in Pleiner et al, the key steps for generating...: Purified nanobodies are then tagged with a fluorescent dye or HRP via maleimide-labeling. Maleimide-...

...Which Fluorescent Protein Should I Use? Plasmids 101: Green Fluorescent Protein (GFP) Fluorescent Protein...Plasmids 101 has covered GFP and its related fluorescent proteins, which are sometimes used as tags for...LVPRGS Cleaves between Arg and Gly residues PreScission LEVLFQGP Cleaves between the Gln and Gly residues...examples. The tables list more common tags than are described in the post, but have been categorized to help...

...plasmid addition was first described by studies of EcoRI and Bsp6I in Escherichia coli in 1995 when introduction...systems. As with the other toxin-antitoxin systems described above, one part of the pair is more stable than...toxin-antitoxin system discovered. Unlike the systems just described, the hok/sok systems consist of three components...proof of concept, they tested a lux-mediated bioluminescence reporter in vivo in the absence of selective..., Hanna Engelberg-Kulka, and Gad Glaser. "An Escherichia coli chromosomal" addiction module" regulated...

...antibiotic’, very often we immediately imagine a prescription for a bacterial infection. While this post will...that the term ‘antibiotic’ can also be used to describe anti-parasitic drugs and anti-fungal drugs, because...development of resistance include (Figure 1): Over-prescription of antibiotics (for example, giving antibiotics...not finishing the whole course of antibiotics prescribed to them This may seem counter-intuitive considering...The take-home? Always take your medication as prescribed. How do we know which bacteria are resistant ...infection is still effectively treated. We can prescribe antibiotics for the shortest time period possible...

...quenched fluorescent ssRNA reporter is added to the reaction. Cleavage of the quenchable fluorescent RNA by...Cas13 and Cas12a, are combined with different fluorescent reporters for each enzyme. In addition, Csm6,...Csm6 and a Csm6 specific reporter (in the same fluorescent channel as the Cas13 reporter), they could significantly...papilloma virus (HPV) genome, via a crRNA. An ssDNA-fluorescently quenched (FQ) reporter, which will produce a...rate of ~1,250 cuts per second. A quantifiable fluorescent signal designates the presence of your DNA of...

...experience and skills would help you do the job as described. Let’s imagine you’re applying to a job that ... your cover letter, you can expand on that by describing how you developed and used a ddPCR assay to answer...start by looking at the requirements in the job description. For each requirement, ask yourself if you have...exactly what you would want to talk about! You could describe your organizational system, your rationale for...company/organization]. I am a [adjective that describes you] scientist with over [number] years of experience...the university]” Include keywords from the job description to help the hiring manager quickly make the connection...

...variability, and in some cases, counter-selection escape can occur: single crossover colonies can acquire... the toxin itself (leading to counter-selection escape). The rhamnose induction system is well-characterized... flexneri, Enterobacter cloacae, and Serratia marcescens and have created dozens of deletions as well ...Matthew J., et al. "Toxic protein expression in Escherichia coli using a rhamnose-based tightly regulated...enables simplified chromosomal gene replacement in Escherichia coli." Appl. Environ. Microbiol. 81.15 (2015)...Recombineering) system for genome editing in Escherichia coli." Scientific reports 5 (2015): 15096. PubMed...

...difficult and time consuming. GAL4/UAS was first described in 1993 by Norbert Perrimon’s lab and has been...Transparency also allows scientists to easily observe fluorescently labelled proteins and tissues to better assess... into the worm, such as animals that express fluorescent markers (Mello et al., 1991). Genes can also ... genetic modification, such as expression of fluorescent protein or disruption of a gene, can be easily...Hesselson, Daniel, et al. "Distinct populations of quiescent and proliferative pancreatic β-cells identified...Read our quick guide to Drosophila Download the fluorescent proteins 101 eBook Read our plasmids 101 blog...

...organisms!) Escherichia coli One of the most recognizable and utilized model organisms is Escherichia coli. ...2017). Escherichia coli as a Model Organism and Its Application in Biotechnology. In Escherichia coli -...environments (Idalia & Bernardo, 2017).  Escherichia coli. Image from NIAID. E. coli is often a...and a tool-kit from the Hamdoun Lab to create fluorescent fusions in a multitude of organisms including...

...antibody worked under the conditions described above, and a short description of how the antibody performed....the submitted application, along with a brief description of the experiment. If that data was included ...ELISA — but in all cases, it contains a detailed description of the experimental conditions.     Figure 4...The submission process is easy, with simple, descriptive fields, drop-down boxes when appropriate, and...

...26322837. 2. Stadler, Charlotte, et al. "Immunofluorescence and fluorescent-protein tagging show high correlation...First described in the 1980s, protein tags are now one of the most useful items in a scientist’s toolbox...displayed good binding efficiency in both immunofluorescence and immunoprecipitation applications, but...Plasmids Tag endogenous proteins with CRISPR Fluorescently Tag Your Protein of Interest ...

...from the event can be found below along with a description of the program beneath it. You can jump to the...the different time stamps in the description to watch any sections you’re particularly interested in, ...antibodies while FPbase curates information about fluorescent proteins so you can best apply these technologies...JoVe, and Bio-protocol all work to provide more descriptive and interactive protocols that make it much easier...Minisymposium on Reproducibility Booklet - Including descriptions of many organizations promoting reproducibility...Addgene Blog FPbase: A new community-editable fluorescent protein database ReFigure: Save Scientific Figures...

... Y, Lacher NA, Ruesch MN. Capillary gel electrophoresis with laser-induced fluorescence of plasmid DNA...confirmations like linear and nicked plasmids, as described in this article.   Long-read sequencing Addgene...; Cook et al., 2020). CGE uses laser-induced fluorescence (CGE-LIF) for detection and can provide data...bacterial colonies using one of the detection methods described above can help ensure the isolation of the monomeric... harboring large inserts are often dimers in Escherichia coli K-12. Plasmid. 1989 Mar;21(2):138-41. doi...

...experience. Most Addgene PhD level job descriptions clearly describe that the roles do not involve bench ...submission of a job application. 3) Read job descriptions carefully Be open minded and be bold. You don...qualifications to apply for a position.  A job description is usually a wish list and the hiring manager... way to a hiring manager. Learn to confidently describe your strengths in your resume and don't be shy...

...resistance (Chen et al., 2013), fluorescence (GFP; Tzur et al., 2013), or rescue of mutant phenotypes (unc-... of lethal mutation co-selection. Only animals rescued for the conditional lethal mutation survive the...experiment, five knock-ins were recovered from seven rescued F1 animals (gel image under worm). Image courtesy...on a plate, other than the parental animal, are rescued progeny, which makes screening extremely rapid....available through Addgene. In 2019, Farboud et al. described another similar co-conversion marker (repair of...

...Runne-Janczy, a Product Development Manager at eScience Labs, an educational company that creates hands-on...scicomm and part two focusing on what her job at eScience Labs is like. Find all the posts in this series...don’t really know where I came from.” Working at eScience Labs, an educational company that creates hands-on...editing and searching for a job and I stumbled upon eScience by luck, by chance, by complete happenstance. ...d had at Iowa really opened the door for me at eScience.” Stay tuned for part two of Caitlin’s interview...

...transduced using only protein. Read on for a description of the lab’s iTOP protein-only transduction method...transduction by osmocytosis and propanebetaine) to describe this method of protein transduction. Later studies...Cre- or Cas9-mediated recombination. Both primary hESCs and immortalized cells treated with iTOP efficiently...populations. After iTOP treatment, both primary (hESCs) and cultured cells showed up to 70% resistance ... frequency of biallelic mutations. Importantly, hESCs retained their stemness, indicating that iTOP may...