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  1. Tips for Writing a Good Cover Letter

    Type
    Blog Post
    ...excellent communication skills” try giving an example, like “I have extensive communication experience...usually yes! A good cover letter won’t hurt your application, but it can certainly help you stand out.  In...letter is vital, particularly in fields where communication and writing are key skills. Other fields, often...consider cover letters an important part of the application. Even within an organization, there can be variance...good cover letter!) As with any part of the job application process, it's a good idea to talk to people in... to proactively address questions about your application. These could be logistical, like briefly explaining...explaining a gap in your job history or sharing relocation plans. They can be skills-based: if you're switching...
  2. Finding nucleic acids with SHERLOCK and DETECTR

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    Blog Post
    ...activated” Cas13 produces a quantifiable signal that indicates the presence of your targeted nucleic acid. To...amplified using RPA (recombinase polymerase amplification) or reverse transcriptase (RT)-RPA, respectively...RNA for subsequent detection by Cas13. This amplification step in combination with the ssRNA reporter ... introduced in 2017 so the lab made further modifications to improve their detection system. They named... SHERLOCKv2 uses far less primer in the pre-amplification step allowing for greater quantitation without...hour. Find the SHERLOCK plasmids at Addgene! Applications of the SHERLOCK detection system The Zhang lab...portability. This method can be used for an array of applications including clinical diagnostics (e.g. pathogen...
  3. Overcoming the AAV Size Limitation for CRISPR Delivery

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    Blog Post
    ...described above showed successful target modification, indicating that AAV is a good delivery vehicle for... with AAV. This system fuses catalytically dead dCas13b to the catalytic domain of RNA deaminase ADAR2...overall packaging capacity but necessitating purification and co-infection of two AAVs. Cas9 orthologs...Streptococcus thermophilus and a rationally-designed truncated Cas9. Unfortunately, certain drawbacks limit the...that limits the number of targetable loci, and truncated Cas9 has much lower efficiency than its wild-type...the AAV-SaCas9-sgRNA constructs mediate genome modification, but they did so without a substantial immune...ADAR2. Constructs containing the ADAR2 truncation ADAR2DD(delta984-1090) are approximately 4.1 kb in length...
  4. CRISPR 101: Targeting RNA with Cas13a (C2c2)

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    Blog Post
    ...use, functional in most species, and has many applications. That said, SpCas9 is not the only game in town... makes this molecule unique, and the various applications of Cas13a. The origins of Cas13a: An RNA cleaving...53 potential candidate genes that fell into 3 categories based on the architecture of the CRISPR protein... is analogous to the PAM sequence for Cas9, is located at the 3’ end of the spacer sequence and consists...discussed below. Subscribe to CRISPR Blog Posts Applications of Cas13a   Figure 1: Using Cas13a as...) is amplified using Recombinase Polymerase Amplification (RPA) or Reverse-transcription RPA (RT-RPA),...fluorophore. The fluorescent signal can be used as an indicator to determine whether the target sequence is present...
  5. Early Career Researcher Toolbox: Free Tools for Making Scientific Graphics

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    Blog Post
    ...software that let me create graphics that better communicated the science in my blog posts. This post highlights...free tools which will hopefully also help you communicate your science, whether it’s in presentations, ...number of drawing tools, so it’s harder to draw intricate figures, like a brain or a mouse. Vectr Vectr ...without the watermark, which are better suited for publications, if you sign up for a paid account.  I’m proud... free medical images organized into four main categories: anatomy and the human body, cellular biology...appropriate credit, provide a link to the license, and indicate if changes were made to the images. This requirement...easier to give attribution, than for journal publications. Google Image Search Google image search is ...
  6. Viral Vectors 101: Parts of the AAV Packaging Plasmid

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    Blog Post
    ...Rep78/68 play a key role in AAV genome replication. Replication initiates from the AAV genome ITRs on ...contains the proteins needed for the virus to replicate,  and the transfer plasmid which contains the ...aka the RepCap plasmid encodes two genes: the replication (rep) gene and the capsid (cap) gene. In wild-type...its products. Created with BioRender.com   Replication (rep) gene The rep gene encodes four proteins...proteins that are required for viral genome replication and packaging: Rep78, Rep68, Rep52, and Rep40. Rep gene... generate a 3’ hydroxyl group needed for DNA replication, Rep78/68 binds the Rep binding element (RBE)...nicked by its endonuclease domain. Following replication, Rep78/68 stays attached to the AAV genome and...
  7. Summer SciComm: Science Storytelling

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    Blog Post
    ...formal scientific communications, they are fantastic fodder for science communication. The trick is to ... is far from the only way to communication science. Science communicators also use the hero’s journey,...this small glimpse into the world of science communication and can use what you've learned to share your...Finding Conflict in Science  In all science communication, what you are doing is telling a (nonfiction...details often aren’t included in scientific communications, but they’re firmly part of the story to the...conflict gives you an easy framework with which to communicate your science. The act of building towards your...your results right at the beginning of your communication, use conflict to get your audience invested ...
  8. Antibodies 101: Normalization and Loading Controls for Western Blots

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    Blog Post
    ...normalization and loading controls needed for relative quantification of a western blot — and why you might want ...accurately compare different lanes? In relative quantification, the POI bands are normalized to the loading...other errors in experimental design and gel quantification. Figure 3: In a perfect world, protein...and it’s also problematic for computational quantification. Outside the linear detection range, band intensity...common error associated with Western blotting quantification” (Gilda & Gomes, 2013). To address this source... Gilda & Gomes, 2013. Figure 7: Semi-quantification of liver lysate using different loading controls...concentration of β-actin in panel A. Indeed, the quantification in panel B is not especially linear. In panel...
  9. Optimizing Donor DNA for Enhanced CRISPR Genome Editing

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    Blog Post
    ... reagent. This interplay falls into two broad categories and thus, causes two types of editing outcomes...altering its function). HDR is crucial for certain applications, for example, correcting the allele that causes... dissociation rates and discovered that the catalytically active form of the nuclease dissociates very...we measured identical dissociation rates for catalytically inactive dCas9 molecules. We followed these ...Our in vitro results also demonstrated that catalytically inactive dCas9 produced a “bubble” structure...nucleofection The in vivo editing experiments in this publication were performed using a technique called nucleofection...Additonal Corn Lab Members who worked on this publication:         References 1. Richardson, Christopher...
  10. Synthetic Photobiology: Optogenetics for E. coli

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    Blog Post
    ...consistent, and controllable pulses of light. Complicated patterns of light intensity and wavelength can... also be used to engineer new pathways with sophisticated gene expression controls. Most cell types do...light; these two systems can be coexpressed for sophisticated control of gene expression. An E. coli light-sensitive...; only two plasmids are required per system. Applications of these systems The large dynamic range of ...suitable for various scientific or engineering applications.   References Refactoring and optimization ...Oct 11. PubMed. Find the plasmids from this publication at Addgene. Synthetic biology: engineering Escherichia...:441-2. PubMed. Find the plasmids from this publication at Addgene. A synthetic genetic edge detection...
  11. 15 Hot Plasmids from 2017

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    Blog Post
    ... plasmid or group of plasmids' functions and applications. We hope that these articles make it easier ...FLIM (Fluorescence Lifetime Imaging Microscopy) applications. To demonstrate the power of mCyFP1, the Lin... Coselection efficiency is so high that some applications may permit working with edited “pools” of cells...showed high editing efficiencies with AsCpf1, indicating that ouabain coselection is a robust and flexible...be found at Addgene and in Figure S12 of the publication. Agudelo D, et al. Nature Methods. 2017. PubMed...potential of base editing for precise genome modification. Plasmid ID Plasmid Name Cas9 Variant Speacial...(NGG)  Very low off-target activity; protein purification vector  87439  pCMV-HF-BE3  HF-Cas9 (NGG) ...
  12. Viral Vectors 101: Types of viruses

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    Blog Post
    ...injections, or the location of a specific cell type doesn’t always lend itself to direct application of virus. ...addition of a helper virus or plasmid allows it to replicate episomally, essentially remaining an extrachromosomal...extrachromosomal part of the host genome that can replicate and divide along with the endogenous DNA. AAV also...gamma-retro viruses. This is large enough for many applications, but in some cases multiple AAVs (co-infection...and AAV. If you’re wondering more about what applications these viruses are good for, we have plenty of...Adenovirus-Mediated Gene Delivery: Potential Applications for Gene and Cell-Based Therapies in the New...Gammaretroviral vectors: biology, technology and application. Viruses. 2011 Jun;3(6):677-713. DOI: 10.3390...
  13. A Deep Dive into BioBuilder

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    Blog Post
    ...and consultant whose work is focused on education and education policy. Additional Resources on the Addgene...that routinely hit the inbox at the BioBuilder Educational Foundation. With a goal of “bringing tomorrow...BioBuilder answers the call by offering a variety of educational programs all focused on current questions and...effectively contribute as part of a team.” As a seasoned educator, Kuldell also knew that the best days in the classroom...As a result, Kuldell launched the BioBuilder Educational Foundation in partnership with high school teachers...grown into a recognized name in the field of STEM education worldwide. The BioBuilder curriculum is now taught...critical thinking, persistence, teamwork, and communication. And as a program focused on an emerging scientific...
  14. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

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    Blog Post
    ...biological pathways;  to observe a phenotype, the modification of multiple genes is often necessary. Guide ...recognition and cleavage. This sequence flexibility indicates that gRNAs could be incorporated into these constructs...the 5’ end. PTG construct schematics. Scissors indicate cleavage sites. After confirming gRNA cleavage...) How many loci can Cas9 modify? To test the application of this system to gRNA multiplexing, Xie et al...deletion with 4-20% frequency at the four loci, indicating that Cas9 can be guided to at least 8 distinct...multiplexing further illustrates the potential of PTGs. Applications of PTGs The major advantage of the PTG system... Mar 2. PubMed. Find the plasmids from this publication at Addgene Other Techniques for gRNA Multiplexing...
  15. Dos & Don'ts When Publishing a Scientific Manuscript

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    Blog Post
    ...should I choose? Writing a research paper for publication in a scientific journal requires several considerations...detailed information on lab resources, such as catalogue numbers and manufacturers’ details for antibodies...Information Framework (NIF) called the Resource Identification Initiative. During this project, authors are...likely that researchers have enough information to replicate your experiments, making research more reproducible...the reproducibility and reliability of science publications. Peer reviewing To make sure your manuscript...happens before the manuscript is accepted for publication and is, in most cases, blind (meaning you don...revised before it can be published, this can delay publication and manuscripts may go through several rounds...
  16. Hot Plasmids: Spring 2025

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    Blog Post
    ...Here's what you'll find in this post: Glutamate indicators with increased sensitivity and tailored deactivation...viral preps targeting interneurons Glutamate indicators with increased sensitivity and tailored deactivation...popular choice for a variety of synaptic imaging applications. Find iGluSnFR4 plasmids here! Aggarwal, A...., Iyer, R., … Podgorski, K. (2025). Glutamate indicators with increased sensitivity and tailored deactivation...DNA vector that contains everything needed to replicate inside a host cell but with space to insert your...This requires both targeting a specific genome location and isolating the interacting proteins. Lucky ...immunoprecipitation. The top panel depicts the identification of heat-shock proteins at the FOS gene promoter...
  17. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

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    Blog Post
    ... Haoyi Qiu He Tian Every two years, we seek applications for awards to early career scientists using ...virtual conference attendance. We also opened up applications to research technicians and undergraduate researchers...mercaptopyruvate sulfurtransferase (MPST) which catalyzes the desulfuration of 3-mercaptopyruvate and generates...the chromophore that are susceptible to redox modification, the MPST hydrosulfide can be transferred to... and CNO silencing. (D) Immunohistochemistry verification of the dual virus expression in CR neurons of...where she engineers genetically encoded voltage indicators. Tian developed a video-based high-throughput...archaerhodopsin‑derived genetically encoded voltage indicator which had improved performance in live mouse brains...
  18. RaPID Detection of RNA-protein Interactions

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    Blog Post
    ...methods Not in a native context. While native purification preserves RNA structure and naturally occurring...used the Contamination Repository for Affinity Purification (CRAPome), to identify highly biotinylated background...research groups that performed BirA*-based affinity purification mass spec experiments. These background proteins...subtracted from Ramanathan et al.’s RaPID datasets. Applications of RaPID RaPID sounds like a nifty technique...help answer? Ramanathan et al. outline three applications of RaPID: Discovery. To discover novel RNA-... that QKI is a host protein critical to Zika replication. Human genetic disease. RaPID can also be used...binding patterns. Hereditary Hyperferritinemia-Cataract Syndrome (HHCS), a disease characterized by increased...
  19. Summer Fun at Addgene!

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    Blog Post
    ...Summer Science Communication Series. We’ll be covering the basics of science communication, tips and tricks...science with others, and introducing science communication as a research field. And there will even be ...be a few tips for scientific communication (manuscripts, talks, etc…!)  Deep Dives You probably are familiar... Dives” series comes in! We’ll take a common application or concept and go beyond the basics - diving ...you’re ready for the next steps on some common applications, join us as we get into the weeds on some of... the end of this month! We’ll be sharing our educational resources and tips and tricks for those new to...media endeavors - in between some much-deserved vacations, sunshine, and beach trips, of course.    *not...
  20. Plasmids 101: The Wide World of Natural Plasmids

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    Blog Post
    ...Shareck et al., 2004). Plasmid categorization is complex Plasmid classification has shifted over the years...us! A plasmid is a small piece of DNA that can replicate independently from its host’s chromosome. Although...genes between bacteria can also have serious implications for public health. Bacteriocin plasmids: microbial...poisoning, the difference is virulence plasmids! This category includes genes that help bacteria invade hosts...years, and new classifications are still being created as whole-genome sequencing becomes more relevant (... common plasmid features, such as origins of replication and promoters, check out our other Plasmids 101...Redondo-Salvo, S., & de la Cruz, F. (2023). Plasmid classifications. Plasmid, 126, 102684. https://doi.org/10.1016...
  21. Early Career Researcher Toolbox: Finding Relevant Papers

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    Blog Post
    ...show all publications by an author, total number of citations, and the number of publications that heavily...read the abstract or full text of the paper, publication date, a relevance score, and type of article....that term. You can also set up alerts for new publications from a particular author. Saving papers Papers... work, so it’s also a good place to look for publications. Bonus: papers have to be summarized in 280 ...organizations, and universities often tweet about publications in the fields of research they support. Monitoring...Artificial Intelligence. Semantic Scholar analyzes publications and figures using machine learning to extract...your search to a specific window of time for publication. To help you quickly identify papers with greater...
  22. Adapting Toehold Switches to Detect Zika Virus

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    Blog Post
    ... journal Cell just six weeks later. The Zika publication timeline I think we can speak for all the authors...research, but the urgent need for the test did catalyze our resolve to get the work done as quickly as... dozen versions of the toehold switches and amplification primers. This helped make sure that we had a...this includes primer directed isothermal RNA amplification, new toehold switch designs, CRISPR/Cas9-mediated...our electronic reader that provides portable quantification of diagnostic results.  Our approach now detects...directly from infected monkey blood plasma without purification or concentration of the virus. Future work Going...focused on developing cell-free synthetic biology applications for human health and the environment.     Alexander...
  23. Illuminating Epigenetics with A FRET Based Biosensor

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    Blog Post
    ...Mechanisms of epigenetic modifications The main mechanism of epigenetic modification is chromatin remodeling...literally means ‘on top of the genome’). Epigenetic modifications can result in phenotypic changes without any...any change in genotype. Epigenetic modifications play a large role in turning genes ‘off’ and ‘on’, signaling...two main mechanisms, either through histone modifications, which can influence the transcription of genes...hypomethylation, on the other hand, has also been implicated in the development and progression of cancer ...enzyme digestion, DNA adenine methyltransferase identification, and bisulfite sequencing. However none of ...can be used for in-situ research of dynamic modifications in real-time, posing a huge challenge to studying...
  24. The Materials Science of Optogenetics Experiments

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    Blog Post
    ...There are several important considerations with fabrication of the fiber optics needed for optogenetics. ... for the implant depends on the dorsal/ventral location of your region of stimulation (the z-axis bregma...emitted from the end of the fiber optic implant scatters a great deal once it enters tissue, placement ...0.5mm above the site of your stimulation. The scatter effect is also an important consideration when ...Lasers can be purchased through Laserglow, which caters to optogenetics users and adjusts the lasers appropriately...require stimulation only under certain contexts/locations in the test apparatus and thus individual laser...inactivation kinetics. For example, the light-gated cation channel, Channelrhodopsin 2 (ChR2), exists in several...
  25. A Guide to Getting Started in Undergrad Research

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    Blog Post
    ...entire page dedicated to different career options from science policy to science communication and much ...), about their experiences. Research requires dedication and effort, so building a strong understanding...critically about whether it’s something you want to dedicate your time and energy to. How can I make the most... feel less confident in their abilities and qualifications. Studies on imposter syndrome have also found...universities often have scholarships and funds dedicated for undergraduates interested in doing research...the experience would improve my scientific and application writing skills, and ended up with the award. ...fantastic for broadening your experiences! These applications can be highly competitive, so asking for help...
  26. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

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    Blog Post
    ...for successful modifications, efficiency can be low, especially for larger modifications. What’s the solution...several potential CRISPR applications. First, CRISPR can be used to target locations in the genomes of pathogenic...tools available for bacteria and some of the applications for which they’ve been used. The beginnings ... of Enterobacteriaceae, without the need for modification. This finding suggests that the system may be...eukaryotes (Qi et al., 2013). As in other systems, catalytically dead (dCas9) targeted to a promoter or gene ...responsive promoter to drive expression of a catalytically dead Cas9 enzyme allows reversible repression...Bikard et al., 2013). By targeting either a catalytically dead Cas9 (dCas9) alone or a dCas9 fused to ...
  27. Lentiviral Vector Uses and Overview

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    Blog Post
    ...many CRISPR applications, and they’ve also had success in clinical gene therapy applications. Read on to...to learn more about the current (and future) applications of lentiviral vectors! Lentiviral vector history...original system, the possibility of creating a replication-competent virus via recombination between the...of recombination events necessary to create a replication-competent virus. Second, Tat is eliminated through...barcoding libraries have been used for complex applications like lineage tracing in heterogeneous tumors...2015: lentiCRISPR v2, used both for targeted modification and genome-wide screening.   Lentiviral vectors...developed integrase-deficient lentiviral vectors for applications that require the large genetic capacity of a...
  28. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

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    Blog Post
    ...manuscript to get reviewed and edited prior to publication, and this will delay the dissemination of your...deposited by George Church’s lab). Addgene has also cataloged a variety of useful molecular cloning protocols...bacterial transformation. And more plasmid-related educational resources can be found in our Plasmids 101 blog...bring your phone into the lab? Maybe having a dedicated lab-use tablet? The software also makes it easy...for biologists and bioengineers to share their educational materials, protocols, and courses, as well as...equipment, and track inventory and orders. Quartzy caters to life science labs and offers the service free...scientists by allowing companies to pay publish their catalogs on the site. Like Labguru, Quartzy offers software...
  29. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

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    Blog Post
    ...emission wavelengths, which are pivotal in their application in biotechnology (Grimm & Lavis, 2022). These... substrates/peptides and antibodies. Example applications of this include coupling to an antibody, whereby...whereby excitation of blue light will indicate where FITC is bound through green light emission. Key considerations...typically bind to. Stability of fluorescence: For applications that involve ranges of pH, and significant lengths...light, is another factor to consider. If your application requires longer periods of exposure, you’ll want...degrees of β-arrestin-2 recruitment to receptors, indicating their agonistic nature. The potency and efficacy...descriptions, or technical manuals. Specific applications and matching dye/ligand choices For effective...
  30. Viral Vectors 101: Systemic Capsids

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    Blog Post
    ... 2023 * indicates viral preps available. Mechanisms of crossing the BBB are indicated in parentheses...suitability of a systemic capsid for an individual application is dependent on a number of factors, including...test your chosen capsid before using it in an application.     Figure 1: AAV toolkit for targeted...doses are recommended for all systemic capsid applications. For example, AAV-PHP.eB is typically administered...experimentally determine the best titer for your application.   Pro tip!Want to know if a capsid is a good... capsid immunogenicity is a concern, using a purification method that removes, not just reduces, empty...always, validate the capsid for your specific application and animal model.  Other model organisms Tropism...
  31. Using Video to Share Your Science: We Share Science

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    Blog Post
    ...be part of the answer. Problems with written publications Online tools like ResearchGate, GoogleScholar... that the big breakthroughs in my discipline, education, will come from ideas garnered from research in...from the science of others. Improving science communication through video     Video abstracts about research... to be that video is the future in terms of communicating ideas. In 2012, for example, 3 billion hours...scientific videos were for entertainment or basic education (i.e., learn why water freezes) there was a growing...video usurps the written word as the means for communicating research. Short videos of 3 to 5 minutes provide... researchers to leverage the video medium to communicate their research more effectively.” How to share...
  32. Antibodies 101: Reading a Flow Plot

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    Blog Post
    ...done using side scatter (SSC) and forward scatter (FSC), which measure light that “scatters” past the cell... the plot that divides it into four, using the location of the cell populations as a guide  (Figure 3....be used for this purpose.  Pro tip! The forward scatter is a proxy for diameter and volume of the cell,...the exclusion of doublets and debris. The side scatter roughly measures granularity, a proxy for nucleic... will have numbers by gated populations; this indicates the percentage of total events contained in the... gated population. Heat maps are often used to indicate the relative density of cells. You can see both...presented. A horizontal line across the peak will indicate the cut-off value for a positive signal (Figure...
  33. Hot Plasmids - November 2023

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    Blog Post
    ...Figure 1: Choosing the best PE6 variant for your application. Image reused form Doman and Pandey et al., 2023... CreVLPs, and eVLPs for in vivo and in vitro applications.   Figure 2: Modular features of DIRECTED...approach for experimental therapies and medical applications.  Find DIRECTED plasmids here! Strebinger, D...delivery by modular envelope design. Nature Communications, 14(1), 5141. https://doi.org/10.1038/s41467...interactions or map a local proteome, but some applications have been challenging or inaccessible. The Ting... (which had therefore been trafficked between locations). Figure 3: A) LOV-Turbo becomes active...active when illuminated with blue light, catalyzing the biotinylation of nearby proteins. B) TransitID uses...
  34. Plasmids 101: E. coli Strains for Protein Expression

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    Blog Post
    ...expression vector cannot have p15A origin of replication F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) pLysS...expression vector cannot have p15A origin of replication F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) pLysE...Expression vector cannot have p15A origin of replication F- ompT hsdSB(rB- mB-) gal dcm (DE3) pRARE2 (...Expression vector cannot have p15A origin of replication fhuA2 [lon] ompT gal (λ DE3) [dcm] ∆hsdS/ pLemo...Expression vector cannot have p15A origin of replication F-, Φ80ΔlacM15, thi, lac-, mtl-, recA+, KmR Expression... commercial strains carry the DE3 lysogen, as indicated by the name of the strain. Conversely, other strains... for positive selection and a p15A origin of replication, making it incompatible with other p15A plasmids...
  35. Increasing Your Work's Visibility with Addgene: Citation, Search, and Collections

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    Blog Post
    ...Addgene ID number, your publication will be listed on our website as a “Publication Citing this Plasmid” ...upon other researchers being able to find their publications in the first place. As a curator of the thousands...make it easier for you to find the plasmids and publications relevant to your research interests. We do this... this by promoting citation, supporting sophisticated search, and curating plasmid collections. Addgene's...number as well as the reference for its original publication. To help facilitate this process, we explain ...fruits of your labor and won’t have to repeat the modification. This will save both time and money. A quick...whether the plasmid has been used in your system. Sophisticated search You can search for specific plasmids ...
  36. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

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    Blog Post
    ... use the GAL4-UAS system along with precisely truncated attP sites to control recombination of the gene...of PhiC31. In the SPARC system, attP sites are located on either side of a stop cassette, upstream of ...amount of labeling, you can alter the attP site. Truncating the attP site upstream of the stop cassette results...population is directly correlated to the length of the truncated attP site. The three SPARC variants, based on ...intermediate (d), and sparse (e) labeling of the calcium indicator, GCaMP6f, in the Drosophila optic lobe. Image ...Image from Isaacman-Beck et al., 2019. Applications for SPARC SPARC technology is versatile and can be ...dendrites of T5 neurons by placing the calcium indicator GCaMP6f in the effector position, as well as to...
  37. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

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    Blog Post
    ...study its function is one of the most popular applications of fluorescent proteins. These fusion proteins...steric hindrance. For instance, GFP may occlude catalytic sites or obstruct binding sites or motifs that...that are necessary for post-translational modifications. Despite these potential issues, GFP has been used...linker sequence short In the early days of GFP applications, many were concerned with steric hindrance by...N-terminus of the donor fluorescent protein can be truncated to increase the FRET efficiency. Creating fusions...fluorescent protein and protein of interest (POI). To indicate the sequence, people often use terminology like...  Determining other aspects of functionality (catalytic activity or interactions) can be challenging. ...
  38. Multiple Plasmids at a Low Price: Kits from Addgene

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    Blog Post
    ...well as protease sites to facilitate protein purification. We hope this kit will allow you to quickly ...for a variety of synthetic biology or protein purification studies in E. coli. 800 and 1900 Series If you...polycistronic E. coli expression constructs. Some applications of plasmids from this kit include the production...and Charles E. Melançon III. "Detection and Quantification of Ribosome Inhibition by Aminoglycoside Antibiotics...designed the pCri system to simplify the protein purification process by making it easy for a researcher to... blog post on the system Easing the Protein Purification Process with pCri Cell-free protein expression...Daniel, et al. "A pull-down procedure for the identification of unknown GEFs for small GTPases." Small GTPases...
  39. Using Phosphoserine to Study Protein Phosphorylation

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    Blog Post
    ...most inclusive databases of post-translational modifications, identifies a staggering ~250,000 phosphorylation...steric and charge-based nature of the phosphoryl-modification; they are both much smaller, and carry only ...thus cannot reflect the true state of protein modification within the context of its appropriate regulation...codons (TAG), replacing them with TAA (7). This modification allowed the deletion of release factor 1, the...optimize protein production using standard protein purification strategies; expression should be optimized for...-incorporated to non-phosphorylated protein. Verification of site-specific pSer incorporation can also...programmable protein phosphorylation." Nature communications 6 (2015). PubMed PMID: 26350500. PubMed Central...
  40. What the HEK?

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    Blog Post
    ... This allows them to replicate viral plasmids with the SV40 origin of replication. To generate a virus...isolating the cell line all the way to modern applications. What the HEK is it?   HEK cells were first...cell biology, there are thankfully many other applications that they are useful for! What the HEK is it...cell line and we will highlight some of those applications here.   Virus production HEKs are the go-to ...proteins at high levels. In a case where clinical applications of recombinant proteins are necessary, HEKs ...significantly reduces the cost of production. Research applications There are two areas of research where HEK cells...vulnerable to viral infection. Yes, your cells can catch a cold—or worse! HEK-293T and its derivatives are...
Showing: 321 - 360 of 1023 results